Mammals have 3 pairs of major salivary glands i.e., the parotid, submandibular, and sublingual glands. Saliva secretion of these glands is modulated by taste perception. Salivary glands are composed mainly of acinar and ductal cells. Primary saliva is secreted by acinar cells and modified during ductal flow. Recently, of the murine 35 bitter taste receptors, Tas2r108 was expressed at highest levels in the submandibular gland by qPCR. Further, Tas2r108-transfected cells respond to a range of bitter compounds, such as denatonium, quinine, colchicine, diphenidol, caffeine and dapson. The objective of the present study was to characterize the expression of Tas2r108 mRNA in acinar and/or ductal cells of the submandibular gland using in situ hybridization (ISH). Male 42-60 days old DBA2 mice were used in the study. Messenger RNAs were extracted from the submandibular gland for generating digoxigenin (DIG) labeled-cRNA probes. These probes were transcribed in anti-sense and sense orientation using T7 RNA polymerase. Dot blot hybridization was performed using DIG labeled-cRNA probes, in order to estimate integrity and optimal diluting concentration of these probes. Subsequently, ISH was performed on murine submandibular gland to detect Tas2r108 mRNA. Dot blot hybridization data demonstrated that Tas2r108 DIG labeled-cRNA anti-sense probes specifically detected Tas2r108 cDNA. ISH results showed that the anti-sense probes labeled acinar and ductal cells in the submandibular gland, whereas no staining was visible in sense controls. Interestingly, the Tas2r108 expression levels were higher in acinar than ductal cells. These results suggested that Tas2r108 might be more associated with primary saliva secretion than with ductal modification of saliva composition.

Temperature as a major environmental factor affects on organisms on various levels including molecular, physiological, behavioral and ecological levels. Transient receptor potential channels (TRPs) are a cation channel family. Among them, thermo-TRPs are known as a thermosensor. The potential role of thermo-TRPs have been identified in the fruit fly, Drosophila melanogaster, in thermotaxis and in thermal acclimation. With RNA interference (RNAi) technique, the role of thermo-TRPs in the red flour beetle, Tribolium castaneum, was identified by measuring thermal avoidance behavior in a behavioral assay. RNAi of trpA1 reduced high temperature avoidance, 39 and 42 °C. Moreover, the effects of RNAi of thermo-TRPs on the heat-induced knockout and the death after short exposure to high temperature was measured after one minute exposure at 52 °C, either with or without a 42 °C 10-minute thermal acclimation period. Even though it was relatively short time exposure to high temperature, it was enough to induce high temperature thermal acclimation. RNAi of trpA1 made faster knockout at 52 °C. With RNAi of painless, the recovery rates from heat-induced knockout after thermal acclimation. RNAi of pyrexia reduced long-term total survivorship without thermal acclimation.

Antennal olfactory receptor neurons (ORNs) for pheromone and plant volatile compounds were identified and characterized in Sitona lepidus and S. discoideus, using the single sensillum recording technique with five pheromone-related compounds and 40 host and non-host plant volatile compounds. Different types of olfactory sensilla containing specialized ORNs were identified in these weevils. Different types of sensilla housed ORNs specialized for pheromone-related compounds, 4-methyl-3,5-heptanedione or one or more of four stereoisomers of 5-hydroxy-4-methyl-3-heptanone. In addition to the pheromone-related ORNs, several types of olfactory sensilla contained ORNs responsive to plant volatile compounds. Most of the ORNs showed high specificity to specific volatile compounds although some of the active compounds showed overlapping response spectra in the ORNs across different types of sensilla. Our study indicates that both S. lepidus and S. discoideus have species-specific set of highly sensitive and selective ORNs for pheromone and plant volatile compounds. The behavioral implication of these findings is discussed.

Chronic/cyclic neutropenia, leukocyte adhesion deficiency syndrome, Papillon-Lefèvre syndrome, and Chédiak-Higashi syndrome are associated with severe periodontitis, suggesting the importance of neutrophils in the maintenance of periodontal health. Various Toll-like receptor (TLR) ligands are known to stimulate neutrophil function, including FcR-mediated phagocytosis. In the present study, the effect of TLR2 activation on the non-opsonic phagocytosis of oral bacteria and concomitant production of reactive oxygen species (ROS) by human neutrophils was evaluated. Neutrophils isolated from peripheral blood were incubated with Streptococcus sanguinis or Porphyromonas gingivalis in the presence of various concentrations of Pam3CSK4, a synthetic TLR2 ligand, and analyzed for phagocytosis and ROS production by flow cytometry and chemiluminescence, respectively. Pam3CSK4 significantly increased the phagocytosis of both bacterial species in a dose-dependent manner. However, the enhancing effect was greater for S. sanguinis than for P. gingivalis. Pam3CSK4 alone induced ROS production in neutrophils and also increased concomitant ROS production induced by bacteria. Interestingly, incubation with P. gingivalis and Pam3CSK4 decreased the amounts of ROS, as compared to Pam3CSK4 alone, indicating the possibility that P. gingivalis survives within neutrophils. However, neutrophils efficiently killed phagocytosed bacteria of both species despite the absence of Pam3CSK4. Although P. gingivalis is poorly phagocytosed even by the TLR2-activated neutrophils, TLR2 activation of neutrophils may help to reduce the colonization of P. gingivalis by efficiently eliminating S. sanguinis , an early colonizer, in subgingival biofilm.

Receptor activator of nuclear factor-κB ligand (RANKL) is an osteoblast/stromal cell-derived essential factor for osteoclastogenesis. During endochondral bone formation, hypertrophic chondrocytes calcify cartilage matrix that is subsequently resorbed by osteoclasts in order to be replaced by new bone. Hypoxia-induced upregulation of RANKL expression has been previously demonstrated in an in vitro system using osteoblasts; however, the involved mechanism remains unclear in chondrocytes. In the present study, we investigated whether hypoxia regulates RANKL expression in ATDC5 cells, a murine chondrogenic cell line, and hypoxiainducible factor-1α (HIF-1α) mediates hypoxia-induced RANKL expression by transactivating the RANKL promoter. The expression levels of RANKL mRNA and protein, as well as HIF-1α protein, were significantly increased in ATDC5 cells under hypoxic condition. Constitutively active HIF-1α alone significantly increased the levels of RANKL expression under normoxic conditions, whereas dominant negative HIF-1α reduced hypoxia-induced RANKL expression. HIF-1α increased RANKL promoter reporter activity in a HIF-1α binding element-dependent manner in ATDC5 cells. Hypoxia-induced RANKL levels were much higher in differentiated ATDC5 cells, as compared to proliferating ATDC5 cells. These results suggested that under hypoxic conditions, HIF-1α mediates induction of RANKL expression in chondrocytes; in addition, hypoxia plays a role in osteoclastogenesis during endochondral bone formation, at least in part, through the induction of RANKL expression in hypertrophic chondrocytes.

Nitric Oxide (NO) is an important signaling molecule in the nociceptive process. Our previous study suggested that high concentrations of sodium nitroprusside (SNP), a NO donor, induce a membrane hyperpolarization and outward current through large conductances calcium-activated potassium (BKca) channels in substantia gelatinosa (SG) neurons. In this study, patch clamp recording in spinal slices was used to investigate the sources of Ca²+ that induces Ca²+-activated potassium currents. Application of SNP induced a membrane hyperpolarization, which was significantly inhibited by hemoglobin and 2-(4-carboxyphenyl) -4,4,5,5- tetramethylimidazoline-1-oxyl-3-oxide potassium salt (c-PTIO), NO scavengers. SNP-induced hyperpolarization was decreased in the presence of charybdotoxin, a selective BKCa channel blocker. In addition, SNP-induced response was significantly blocked by pretreatment of thapsigargin which can remove Ca²+ in endoplasmic reticulum, and decreased by pretreatment of dentrolene, a ryanodine receptors (RyR) blocker. These data suggested that NO induces a membrane hyperpolarization through BKca channels, which are activated by intracellular Ca²+ increase via activation of RyR of Ca²+ stores.

Toll-like receptor 7 (TLR7) is critical for the triggering of innate immune response by recognizing the conserved molecular patterns of single-stranded RNA (ssRNA) viruses and mediated antigenic adaptive immunity. To understand how TLR7 distinguish pathogen-derived molecular patterns from the host self, it is essential to be able to identify TLR7 receptor interaction interfaces, such as active sites or R848-agonist binding sites. The functional interfaces of TLR7 can serve as targets for structure-based drug design in studying the TLR7 receptor’s structure-function relationship. In contrast to mammalian TLR7, chicken TLR7 (chTLR7) is unknown for its important biological function. Therefore, it has been targeted to mediate contrasting evolutionary patterns of positive selection into non-synonymous SNPs across eleven species using TLR7 conservation patterns (evolutionary conserved and class-specific trace residues), where protein sequence differences to the TLR7 receptors of interest record mutation that have passed positive section across the species. In this study, we characterized the Lys609 residue on chTLR7-ECD homodimer interfaces to reflect the current tendency of evolving positive selection to be transfer into a stabilization direction of the R848-agonist/ chTLR7-ECDs complex under the phylogenetically variable position across species and we suggest a potential indicator for contrasting evolutionary patterns of both the species TLR-ECDs.

무당벌레(Harmonia axyridis)는 주로 진딧물을 포식하는 진딧물의 주요 천적으로 알려져 있으나, 무당벌레에 관한 유전자 정보에 대해서는 알려진 바가 거의 없다. 기존 연구에서 RNAi(RNA interference)를 이용하여 무당벌레의 유전자 기능 분석을 위한 유전자들을 선발하기 위해 gateway system을 이용하여 무당벌레 cDNA library를 제작하였다. 그 결과 RNAi에 적합한 200bp~400bp의 insert를 확인하였으며, 최종적으로는 2.58×106 titer의 무당벌레 cDNA library를 제작하였다. 라이브러리 임의의 유전자를 클로닝하기 전에 transcription vector를 이용한 클로닝 시스템을 확인하기 위해서 기존의 무당벌레 탈피 관련 수용체인 ecdysone receptor A를 knock-down시켜 탈피가 억제되는지를 확인하였다. 그 결과 dsEcRA를 주입한 4령 무당벌레는 대부분 번데기가 되지 못하고 죽었으며, qRT-PCR한 결과 무처리에 비해 dsEcRA를 주입한 개체의 경우 주입 후 4일차에 발현량이 감소한 것을 확인했다. 기존의 무당벌레 유전자 기능분석을 위해 제작한 무당벌레 cDNA library 임의의 유전자들을 transcription vector인 LITMUS 28i vector에 클로닝 하여 dsRNA를 합성한 뒤 무당벌레에 주입하여 체내에서 표현형의 변이가 나타나는 유전자를 선발한다. 그 후에 race를 통해 유전자의 전체 시퀀스를 알아내 유전자 정보를 확인함으로서, 무당벌레 내 유전자 기능 분석에 도움이 될 수 있을 것으로 사료된다.

Cryopreservation of boar semen is continually researched in reproductive technologies and genetic resource banking in breed conservation. For evaluating the boar semen quality, sperm motility (MOT) is an important parameter because the movement of spermatozoa indicates active metabolism, membrane integrity and fertilizing capacity. Various researches have been trying to improve the quality of semen Post-thawed in boar. Recently, polymorphism (g. 35756 T>C) of Estrogen Receptor 1 (ESR1) gene reported to be significant association with MOT. This study was conducted to evaluate the ESR1 gene as a positional controlling for motility and kinematic characteristics of post-thawed boar semen. To results, The g.35756 T>C SNP of ESR1 was significantly associated with frozen semen motility and kinematic characteristics. The g.35756 T>C SNP was high significantly associated with MOT, VCL, VSL and VAP (p<0.001). The SNP was also significantly associated with ALH (P<0.05). Therefore, we suggest that the g. 35756 T>C polymorphism in the intron 1 region of the porcine ESR1 gene could potentially be applied in frozen semen programs to improve MOT trait, but only after validation in other populations.

Although it is believed that internal nutrient sensors play important roles in feeding behaviors, their molecular and neural mechanisms underlying of the modulation of physiological status and cell growth are poorly understood. Using a Ca2+ imaging experiments with heterologous expression systems, we show that one of the gustatory receptors in the western honey bee Apis mellifera is selectively tuned to amino acids. Remarkably, we report that this gustatory receptor of the honey bee is highly expressed in hypopharyngeal gland, which plays a role in caste differentiation as well as royal jelly production and secretion. Knocking down this gustatory receptor gene reduces cellular pathways responsible for nutritional sensing such as mTOR signals in hypopharageal gland. Furthermore, the interfering expression of this gustatory receptor gene not only alters morphological changes and developmental retardation of the hypopharyngeal gland, but it also blocks cellular growth signals to induce autophagy. This new report indicates that internal sensing and downstream signals detecting nutrients is essential for honey bee to maintain the cellular growth and development of internal organs essential for caste development and maintenance of social structure in the honey bee.

Evolution of resistance to entomopathogenic bacterium, Bacillus thuringiensis (Bt), can potentially reduce the efficacy of insecticidal proteins from Bt to insect pests in fields. Bt resistance is involved in modification of the toxin binding to its specific midgut membrane receptors, such as cadherin, aminopeptidase N, alkaline phosphatase, and ABC transporters. The beet armyworm, Spodoptera exigua, is one of major lepidopteran insect pest in Korea and showed highly susceptible to Cry1Ca. We investigated the Cry1Ca toxicity with respect to its binding affinity to a Bt receptor, cadherin compared with Cry1Ac. RNA interference (RNAi) of a cadherin of S. exigua (SeCad1) significantly suppressed the Cry1Ca to the toxic level of Cry1Ac. Binding affinity of Cry1Ca to brush border membrane vesicle (BBMV) of S. exigua midgut was significantly lost after SeCad1 RNAi. Binding affinity of Cry1Ac to BBMV was much low compared to that of Cry1Ca and less sensitive to SeCad1 RNAi. Direct binding assay of Cry toxins to SeCad1 was assessed using a recombinant cadherin repeat 10-11 (rCR10-11) of SeCad1. The addition to rCR10-11 to Cry1Ca significantly enhanced the toxicity under SeCad1 RNAi. However, the synergistic effect of rCR10-11 on toxicity of Cry1Ac was not much significant with poor binding affinity of Cry1Ac compared to Cry1Ca. These results indicate that the differential toxicity of Cry toxins against S. exigua is caused by the different affinities to the Bt receptor, cadherin.

Osteocytes may function as mechanotransducers by regulating local osteoclastogenesis. Reduced availability of oxygen, i.e. hypoxia, could occur during disuse, bone development, and fracture. Receptor activator of nuclear factor-κB ligand (RANKL) is an osteoblast/stromal cell derived essential factor for osteoclastogenesis. The hypoxia induced osteoclastogenesis via increased RANKL expression in osteoblasts was demonstrated. Hypoxic regulation of gene expression generally involves activation of the hypoxia-inducible factor (HIF) transcription pathway. In the present study, we investigated whether hypoxia regulates RANKL expression in murine osteocytes and HIF-1α mediates hypoxia-induced RANKL expression by transactivating RANKL promoter, to elucidate the role of osteocyte in osteoclastogenesis in the context of hypoxic condition. The expression levels of RANKL mRNA and protein, as well as hypoxia inducible factor-1α (HIF-1α) protein, were significantly increased in hypoxic condition in MLO-Y4s. Constitutively active HIF-1α alone significantly increased the levels of RANKL expression in MLO-Y4s under normoxic conditions, whereas dominant negative HIF-1α blocked hypoxia-induced RANKL expression. To further explore to find if HIF-1α directly regulates RANKL transcription, a luciferase reporter assay was conducted. Hypoxia significantly increased RANKL promoter activity, whereas mutations of putative HIF-1α binding elements in RANKL promoter prevented this hypoxia-induced RANKL promoter activity in MLO-Y4s. These results suggest that HIF-1α mediates hypoxia-induced up-regulation of RANKL expression, and that in osteocytes of mechanically unloaded bone, hypoxia enhances osteoclastogenesis, at least in part, via an increased RANKL expression in osteocytes.

Six of cadherins have been selected from the P. xylostella genome 52 open reading frames are annotated as cadherin-like genes. Compared to other 5 cadherins of P. xylostella (PxCads), PxCad1 has the highest homology with other lepidopteran insect cadherins and PxCad1 was expressed in all developmental stages specially in gut tissue. Expression of PxCad1 was suppressed by feeding its specific double-stranded RNA (dsRNA, 150ng/larva) and treatment of dsPxCad1 significantly reduced susceptibility to Bt Cry1Ac toxin. To confirm the specific interaction between PxCad1 and Cry1Ac, a toxin-binding assay was performed using enzyme-linked immunosorbent assay (ELISA). The ELISA indicates that BBMV extracted from PxCad1-silenced P. xylostella have significantly lower binding activity to active form of Cry1Ac than control BBMV. Moreover, the analysis of the binding parameters showed that the toxin affinity (Kd) of the control BBMV extract (BBMV-dsCON) was 6.08 ± 0.84 nM, which was not much different to the affinity value (6.72 ± 0.81 nM) of the dsPxCad1 treatment. However, there was a remarkable difference in number of binding sites (Bmax), in which BBMV-dsCON extract had 1.61 ± 0.04, but the BBMV-dsPxCad1 extract had 0.88 ± 0.02. Taken together, these results are suggest that PxCad1 is a functional receptor for Cry1Ac toxicity against P. xylostella larva.

Phytophagous insects detect volatile compounds produced by host and non-host plants, using species-specific sets of olfactory receptor neurons (ORNs). To investigate the relationship between the range of host plants and the profile of ORNs, single cell recordings were carried out to identify ORNs and corresponding active compounds in female Uraba lugens (Lepidoptera: Nolidae), an oligophagous eucalypt feeder. Based on the response profiles to 39 plant volatile compounds, 13 classes of sensilla containing 40 classes of ORNs were identified in female U. lugens. More than 95% (163 out of 171) of these sensilla contained 16 classes of ORNs with narrow response spectra, and 62.6% (107 out of 171) 18 classes of ORNs with broad response spectra. Among the specialized ORNs, seven classes of ORNs exhibited high specificity to 1,8-cineole, (±)-citronellal, myrcene, (±)-linalool and (E)-β-caryophyllene, major volatiles produced by eucalypts, while nine other classes of ORNs showed highly specialized responses to green leaf volatiles, germacrene D, (E)-β)-farnesene and geranyl acetate that are not produced by most eucalypts. We hypothesize that female U. lugens can recognize their host plants by detecting key host volatile compounds, using a set of ORNs tuned to host volatiles, and discriminate them from non-host plants using another set of ORNs specialized for non-host volatiles. The ORNs with broad response spectra may enhance the discrimination between host and non-host plants by adding moderately selective sensitivity. Based on our finding, it is suggested that phytophagous insects use the combinational input from both host-specific and non-host specific ORNs for locating their host plants, and the electrophysiological characterization of ORN profiles can be used for predicting the range of host plants in phytophagous insects.

The objective of this study was to determine the breed differences in the 3'-untranslated region (UTR) of MC1R mRNA, which may be used to distinguish Korean brindle cattle (Chikso) from other breeds. We investigated the relationship between the variation of 3'-UTR of the MC1R mRNA and coat color among different breeds and the Korean brindle cattle with different coat colors. MC1R mRNA expression levels were determined in accordance with the coat color and hair colors of the tail. Total cellular RNA was extracted from the hair follicles of the tails in Hanwoo, Korean brindle cattle, Holstein and HanwooxHolstein crossbred cattle. After cDNA synthesis, PCR was performed. Sequences of the 3'-UTR of MC1R mRNA were analyzed. The 3'-UTR of the MC1R mRNA from different breeds of cattle did not show any variations. There were no variations in the 3'-UTR of the MC1R mRNA in Korean brindle cattle with different coat colors. The levels of MC1R mRNA expression in hair follicles of the tail varied substantially among the Korean brindle cattle with different coat colors, except yellow coat color. Correlation between the MC1R mRNA expression in the hair follicles of the tail and coat color may be present in the Korean brindle cattle, but not between the variations of 3'-UTR of MC1R mRNA and coat color. Further studies to determine the regulation of MC1R mRNA expression from the hair follicles of different coat colors will be beneficial in clarifying the role of MC1R in the coat colors of the Korean brindle cattle.

This study was conducted for SNPs in the 5'-regions of estrogen receptor-α(ESR-α), and association with calving interval (CI), service per conception (SPC) and 305 days milk yield in Hanwoo and Holstein dairy cattle. The genet-ic improvement was incurred low reproduction performance. The objective of this study was to investigate connec-tions between single nucleotide polymorphisms (SNP) of Estrogen receptor-α (ESR-α) with reproduction performance (calving interval, service per conception, and 305 d milk yield) in Hanwoo and Holstein dairy cattle. Hanwoo and Holstein blood samples were collected from 183 and 124 dam of breeding farms and DNA was extracted. Primer design was based on NCBI GenBank (Accession No. AY340579). The PCR-RFLP method with Bgl I was used to ge-notype the cattle. The result showed two variants of the ESR-α gene. The Bgl I cut the 492 bp amplification pro- duct into 322 bp and 170 bp fragments for allele G, while allele A remained uncut, resulting in two restriction frag-ments for homozygote G/G and three fragments for heterozygote A/G. We found two of different genotypes in the-se breeds, A/G and G/G. In Hanwoo, the A/G genotype frequency was 0.13, and G/G was 0.87. The CI of A/G was 382.18±10.03 days, and G/G was 381.69±5.22 days. The SPC of A/G was 1.62±0.16, and G/G was 1.32±0.04. While CI showed no significance difference, SPC exhibited significant difference (p<0.05). In Holstein cattle, the frequency of genotype A/G was 0.02 and G/G was 0.98. The 305 days milk yield of A/G was 7,253.00±936.00 kg and of G/G was 8,747.51±204.88 kg, showing no significant difference.

Endothelin 2 (EDN2) induces follicular rupture by constricting periovulatory follicles. In this study, it was investigated the mechanisms of EDN2 action on follicular rupture with respect of receptor using the conditionally granulosa cell specific EDN2 receptor type A (ETa) KO mice (gcETaKO; ETaflox/-․Amhr2Cre). It was generated the gcETaKO mice by breeding with ETaflox/- mice after mono-alleic ETa knockout by ZP3Cre and Amhr2Cre mice. Fertility, ovulation and maturation rates of ovulated oocytes after super ovulation were investigated in the gcETaKO mice compared with wild-type mice (ETaflox/flox and ETaflox/-) as a control group. In the gcETaKO mice, normal fertility after breeding with male mice was shown compared with wild-type mice. And, there was no significant differences in ovulation rates after super ovulation, however its maturation rates was lower than that of wild type mice. These findings show that EDN2 in follicular rupture for ovulation is related with an other ETa not in granulosa cells. Further studies are needed to investigate how EDN2 is acted in ovarian follicular rupture for ovulation.

Neuromedin B (NMB) acts as a growth factor or a morphogen and plays a role in cancer progression. Indeed, the NMB receptor (NMB-R) is overexpressed in different types of tumors. In our current study, we investigated the involvement of NMB-R in the proliferation of oral cancer cells. Human oral squamous cell carcinoma (SCC) and human oral cancer cells, SCC-25 cells were found to be NMB-R-positive. The NMB-R antagonist PD168368 inhibited the proliferation of SCC-25 cells and reduced their colony formation capacity. We also found that PD168368 induced the cell cycle arrest and apoptosis of SCC-25 cells in a dose-/time-dependent manner. Overall, this antitumor activity of PD168368 in human oral cancer cells suggests that NMB-R is a potential target for the future prevention and treatment of human cancers.

Brucella spp. are facultative intracellular pathogens that have the ability to survive and multiply in professional and nonprofessional phagocytes and cause abortion in domestic animals and undulant fever in humans. Brucella species can survive in a variety of cells, including macrophages and their virulence and chronic infections are thought to be due to their ability to avoid the killing mechanisms within macrophages. Inhibition of phagosome-lysosome fusion has been proposed as a mechanism for intracellular survival of Brucella in professional and nonprofessional phagocytes. Toll-like receptors (TLRs) are part of a skillful system for detection of invasion by microbial pathogens. Recognition of microbial components by TLRs triggers signaling pathways that promote expression of genes and regulate innate immune responses. Recent studies for the interaction between TLRs-Brucella have indicated the importance of control of Brucella infection. Here, we review selected aspects of TLRs-Brucella interaction, which may be helpful to understanding the mechanism of Brucella pathogenesis.

Toll–interleukin 1 receptor (TIR) superfamily는 intracellular TIR domain에 존 재하며, proinflammatory cytokines을 생산하는 transcription factor NF-κB의 활성 화에 의해 선천성 면역을 개시한다. 본 연구에서는 갈색거저리 유충을 이용하여 Mycoplasma genus와 유사한 acholeplasma lysate를 접종하여 비교 유전체학적 방 법을 통하여 갈색거저리에서 면역반응에 관여하는 t1/st2 receptor binding protein 을 동정하였으며, 그 구조 분석을 위한 기초 데이터를 확보하고자 하였다. IL1RL1 (Interleukin 1 receptor-like 1) gene는 Toll-like receptor superfamily로써 사람에서 발견되며 ST2는 Toll-interleukin 1 receptor family의 member이자 endotoxin tolerance 유지에 중요한 역할을 하는 것으로 알려졌다. 갈색거저리 유충에 acholeplasma lysate를 처리하기 전과 후의 각 샘플들로부터 cDNA library를 구축 한 후 random sequencing 을 통해 분석되어진 서열들 중 증감하는 유전자들을 동정 하였고, 그 중 acholeplasma 처리 후 약 4배 정도 발현이 증가한 t1/st2 receptor binding protein 의 서열을 추출한 후 단백질의 2차 구조를 예측한 결과 alpha helix 구조는 서열상에서 8영역으로 예측되었으며 beta sheet 구조는 서열상에서 1영역 에서 존재하고 있어 후속연구를 진행하고 있다.