Previously, we have performed de novo RNA sequencing of Scolpendra subpinipes mutilans using next generation sequencing technology and identified several AMP candidates. Among them, a synthetic peptide (scolopendrasin I) was designed based on SVM algorithm. In this study, we reported that the synthetic peptide scolopendrasin I had an antimicrobial and anticancer activity. As a result, scolopendrasin I showed antibacterial activities against Gram positive and Gram negative bacteria strains in radial diffusion assay and colony count assay without hemolytic activity. In addition, we confirmed that scolopendrasin I bound to the surface of bacteria via a specific interaction with lipoteichoic acid and lipopolysaccharide, which is one of bacteria cell membrane components. In addition, we found that scolopendrasin I had anticancer activities in the human leukemic T lymphocyte cell line Jurkat using MTS assay. In conclusion, our results suggested that scolopendrasin I could be useful for developing peptide antibiotics and anticancer agents.

To identify genes that are differentially expressed, we compared the mRNA expression profile of Harmonia axyridis larvae untreated and treated with LPS. We extracted mRNAs from the larvae with or without LPS treatment, and subjected them to ACP RT-PCR analysis using a combination of 120 arbitrary primers (ACP1-ACP120)and oligo (dT) primer (dT-ACP2). After synthesized cloning DNA from 37 DEGs, it practiced the sequencing homology analysis using BLAST search. Among the 37 DEGs differentially expressed, we identified a cDNA showing homology with previously reported antimicrobial peptide. A cDNA encoding a 82-mer propeptide was identified and its predicted molecular mass and pI was 9.25 kDa and 7.54, respectively. A 35-mer mature peptide was also selected and named herein as Hamoniasin. The antimicrobial activity of chemically synthesized peptide (Mou def 1~8) against human bacterial pathogens was investigate. the result showed all bacteria strains were susceptible to Mou def 2,8 with MIC values in the 32 uM range. And biological changes of the respective cells according to peptide (Mou def 8) treatment were compared. MTT assay was tested that treatment of Mou def 8 decreased cell viability in AML-2, Jurkat, U937 (maximum 200ug/ml, 24hours). That is, fragmentation of DNA, typical characteristics observed in the process of apoptosis, was confirmed in the nucleus of cells dying owing to Mou def 8 treatment.

This study was performed to comprehend the developmental characteristics of cloned embryos knocked out (KO) of α-1,3-galactosyltransferase (GalT) gene. Immature oocytes were collected and cultured for 40 hrs (1-step) or 20hrs (with hormone) + 20hrs (without hormone) (2-step). The embryos transferred with miniature pig ear fibroblast cell were used as control. The reconstructed embryos were cultured in PZM-3 with 5% CO2 in air at 38.5℃ for 6 days. To determine the quality of the blstocysts, TUNEL and quantitative realtime RT-PCR were performed. The embryos were transferred to a surrogate (Landrace) at an earlier stage of the estrus cycle. The maturation rate was significantly higher in 2-step method than that of 1-step (p<0.05). The blastocyst development of GalT KO embryos was significantly lower than that of normal cloned embryos (p<0.05). The total and apoptotic cell number of GalT KO blastocysts was not different statistically from control. The relative abundance of Bax-α/Bcl-xl ratio was significantly higher in both cloned blastocysts than that of in vivo blastocysts (p<0.05). Taken together, it can be postulated that the lower developmental potential and higher expression of apoptosis related genes in GalT KO SCNT embryos might be a cause of a low efficiency of GalT KO cloned miniature pig production.

Cecropin is an antimicrobial peptide that is synthesized in fat body cells and hemocytes of insect in response to a hypodermic injury or bacterial infection. A 503 bp cDNA encoding a cecropin-like antimicrobial peptide was isolated by employing annealing control primer (ACP)-based differential display PCR and 5'-RACE from immunized Papilio xuthus larvae. The open reading frame (ORF) of isolated cDNA encoded a 63 amino acid prepropeptide with a putative 22-residue signal peptide, a 3-residue propeptide and a 38-residue mature peptide with a theoretical mass of 4060.89 Da. The deduced amino acid sequence of peptide showed significant identities with other Lepidopteran cecropins. This peptide was named as papiliocin. RT-PCR revealed that the papiliocin transcript was detected at significant level after injection with bacterial lipopolysaccharide (LPS). Based on the deduced amino acid sequence of papiliocin, a 38-mer mature peptide was chemically synthesized by Fmoc method, and analyzed antimicrobial activity. The synthetic papiliocin peptide had a broad spectrum of activity against fungi, Gram-positive and negative bacteria, and also showed no hemolytic activity against human red blood cell.

We have previously shown that the larvae of swallowtail butterfly, Papilio xuthus, exhibit substantial antibacterial activity in the hemolymph, upon challenging with bacterial lipopolysaccharide (LPS). Here we report the isolation and molecular characterization of several immune inducible genes that are specifically expressed by employing annealing control primer (ACP)-based GeneFishing polymerase chain reaction (PCR) from P. xuthus the larva. Using 120 arbitrary ACPs, we identified 24 differentially expressed genes (DEGs) that are up-regulated in response to injected LPS. Sequence analysis showed that 18 DEGs revelaed a high sequence similarity to the previously characterized genes of other insects, although 6 DEGs showed no significant similarity to any known genes. Among these inducible transcripts we found 8 putative immune-related genes including cecropin and attacin. Finally, we analysed the expression profiles of potential immune-related genes by RT-PCR and found all of them were considerably increased in the mRNA levels by LPS injection.

One of the reasons to causing blood coagulation in the tissue of xenografted organs was known to incompatibility of the blood coagulation and anti-coagulation regulatory system between TG pigs and primates. Thus, overexpression of human CD73 (hCD73) in the pig endothelial cells is considered as a method to reduce coagulopathy after pig-to-non-humanprimate xenotransplantation. This study was performed to produce and breed transgenic pigs expressing hCD73 for the studies immune rejection responses and could provide a successful application of xenotransplantation. The transgenic cells were constructed an hCD73 expression vector under control porcine Icam2 promoter (pIcam2-hCD73) and established donor cell lines expressing hCD73. The numbers of transferred reconstructed embryos were 127 ± 18.9. The pregnancy and delivery rate of surrogates were 8/18 (44%) and 3/18 (16%). The total number of delivered cloned pigs were 10 (2 alive, 7 mummy, and 1 died after birth). Among them, three live hCD73-pigs were successfully delivered by Caesarean section, but one was dead after birth. The two hCD73 TG cloned pigs had normal reproductive ability. They mated with wild type (WT) MGH (Massachusetts General Hospital) female sows and produced totally 16 piglets. Among them, 5 piglets were identified as hCD73 TG pigs. In conclusion, we successfully generated the hCD73 transgenic cloned pigs and produced their litters by natural mating. It can be possible to use a mate for the production of multiple transgenic pigs such as α-1,3-galactosyltransferase knock-out /hCD46 for xenotransplantation.

Unlike mouse results, cloning efficiency of nuclear transfer from porcine induced pluripotent stem cells (piPSCs) is very low. The present study was performed to investigate the effect of cell cycle inhibitors on the cell cycle synchronization of piPSCs. piPSCs were generated using combination of six human transcriptional factors under stem cell culture condition. To examine the efficiency of cell cycle synchronization, piPSCs were cultured on a matrigel coated plate with stem cell media and they were treated with staurosporine (STA, 20 nM), daidzein (DAI, 100 μM), roscovitine (ROSC, 10 μM), or olomoucine (OLO, 200 μM) for 12 h. Flow Cytometry (FACs) data showed that piPSCs in control were in G1 (37.5±0.2%), S (34.0±0.6%) and G2/M (28.5±0.4%). The proportion of cells at G1 in DAI group was significantly higher than that in control, while STA, ROSC and OLO treatments could not block the cell cycle of piPSCs. Both of viability and apoptosis were affected by STA and ROSC treatment, but there were no significantly differences between control and DAI groups. Real-Time qPCR and FACs results revealed that DAI treatment did not affect the expression of pluripotent gene, Oct4. In case of OLO, it did not affect both of viability and apoptosis, but Oct4 expression was significantly decreased. Our results suggest that DAI could be used for synchronizing piPSCs at G1 stage and has any deleterious effect on survival and pluripotency sustaining of piPSCs.

Abscission is an important developmental process used to shed organs such as leaves, flowers and fruits. Despite the detailed characterization of growth dynamics and hormonal balance during the early steps of fruit development, the molecular aspects remain unclear. Abscission of young fruit occurs by separation of cells in anatomically distinct regions between the pedicel and junction. Differences of gene expression between central pedicel and lateral pedicel were investigated by NGS. Partial cDNAs from 15 clones from both the central pedicel and lateral pedicel were selected for nucleotide sequence determination and homology searches, and 12 clones were subsequently selected for further analysis. In preliminary series of Real Time PCR analysis, 9 genes were confirmed as showing a higher expression level in lateral pedicel than in central pedicel. Many of these genes are expressed in a central or lateral pedicel in specific manner, and the expression profiles of the representative genes were confirmed. To clarify the mechanism of MdIAA14 transcription factor gene underlying abscission zone development, we are investigating the expression patterns between central and lateral pedicels in different apple cultivar using real-time PCR and constructing the vector for transformation into apple.

The Hardy Kiwifruit (A. arguta (Sieb. & Zucc.) Planch. ex Miq.) is one of the valuable species due to their edible fruit, high content of nutritious substances, especially abundant of vitamin C, and distinctive flavor and medicinal usage. In order to develop a new A. arguta cultivar with larger fruit and high yielding, 168 candidate plants were collected from wild population in 12 locations distributed 4 provinces (Chungbuk, Gangwon, Gyeongnam, Jeonbuk) from 1985 to 1987. A clone bank that contained highly productive, superior genotypes of A. arguta was assembled in 1988, and 32 excellent clones were selected in 1996 through the clonal test for the growth and fruiting. Evaluation on these clones was done for the fruiting characteristics, yield trial, and major agronomic traits, such as Average Fruit Length (FL) and Width (FW), and Weight of Fruit (WF) and Individual Yields (IY), during 6 years (1997~2002). After the final selection, a new A. arguta cultivar, “Sae-Han” with larger fruit and high yielding was registered as a new variety denomination and certificated variety production and merchandising in 2006 (Table 1). “Sae-Han” is a tetraploid plant and has ellipsoid fruit shape, and has particularly large fruit size (Fig. 1, Table 3). The major agronomic traits of “Sae-Han” showed the large values and selection effect with an average of 43.6 mm (FL), 36.1 mm (FW), 29.4 g (WF), and 17.5 kg (IY), which are 35.0%, 19.9%, 81.5%, and 124.4% compared to the mean of 29 sample trees, respectively (Table 2).

Hardy Kiwifruit (A. arguta (Sieb. & Zucc.) Planch. ex Miq.) is one of the valuable species due to their edible fruits, high content of nutritious substances, especially abundant of vitamin C, and distinctive flavor and medicinal usage. Therefore, we have developed a new cultivar of A. arguta with large fruits and high yielding. For this, 168 candidate plants were collected from wild population in 12 locations from 4 provinces (Chungbuk, Gangwon, Gyeongnam, and Jeonbuk) from 1985 to 1987. A clone bank that contained highly productive, superior genotypes of A. arguta was assembled in 1988, and 32 excellent clones were selected in 1996 through clone tests for growth and fruiting. From these clones, we have regularly investigated yield trials for the fruiting characteristics for evaluation of major agronomic traits, which are the average of Fruit Length (FL), Fruit Width (FW), Weight of Fruit (WF) and Individual Yield (IY), during consecutive 6 years (1997~2002). Finally, we have selected the new A.arguta cultivar, “Chil-Bo” with large fruits and high yielding and registered as a new variety denomination and certificated for variety production and merchandising in 2006 (Table 1). This cultivar is characterized by a tetraploid plant and spheroid type in the fruit shape, and is particularly characterized by large fruit size (Fig. 1). The major agronomic traits of this cultivar showed the large selection effect with an average of 28.4 mm (FL), 36.9 mm (FW), 18.2 g (WF), and 24.2 kg (IY), which are -12.1%, 22.6%, 12.4%, and 210.3% compared to the mean of 29 sample trees, respectively (Table 2).

“Jokyoung”, a new bread making wheat cultivar, was developed from the cross between “Seri 82”, a hard white wheat from CIMMYT, Mexico and “Keumkang”, a hard white wheat with high milling rate and early maturing from Korea by National Institute of Crop Sci

Molecular markers are useful tools for evaluating genetic diversity and determining cultivar identity. Present study was conducted to evaluate the genetic diversity within a diverse collection of rice accessions used for Korean breeding programs. Two hundred eighty-seven rice cultivars, composed of temperate japonica, tropical japonica, indica, and Tongil-type of Korean crossing parents were evaluated by means of 15 simple sequence repeat (SSR) markers. A total of 99 alleles were detected, and the number of alleles per marker ranged from 4 to 11, with an average of 6.6 per locus. Polymorphism information content (PIC) for each of the SSR markers ranged from 0.2924 to 0.8102 with an average of 0.5785. These results, with the result that use of only 15 SSR markers made all rice cultivars examined could be uniquely distinguished, imply the efficiency of SSR markers for analysis of genetic diversity in rice. Cluster analysis was performed on similar coefficient matrics calculated from SSR markers to generate a dendogram in which two major groups corresponding to japonica (Group I) and indica and Tongil type rice (group II) with additional subclasses within both major groups. The narrowness of the Korean breeding germplasm was revealed by the fact that most of the Korean-bred and Japan-bred temperate japonica cultivars were concentrated into only 2 of the sub-group I-1 (143 cultivars) and I-2 (58 cultivars) among six sub-groups in major group of japonica. This is because of the japonica accessions used in this study was a very closely related ones because of frequent sharing of the crossing parents with similar genetic background with synergy effect of the inherited genetic difference between indica and japonica. A rice breeding strategy with the use of molecular markers was discussed for overcoming of genetic vulnerability owing to this genetic narrowness.

If a quantitative trait loci (QTL) marker identified in a population is applicable to different populations (marker universality), this will not only reduce the labor and cost in marker assisted selection (MAS), but accelerate the application of molecular markers to real breeding programs. Present study aims to evaluate the defined QTL related markers from a population to a different breeding population for the MAS. Four rice breeding populations were subjected to seventy-five simple sequence repeat (SSR) markers which were already identified for their polymorphism information content (PIC) in the parents of the crossings. Among them, eight markers were evaluated for their correlation between presence of marker alleles and phenotypic expression in breeding populations. A reasonable level of polymorphism for the mapped markers originated from any sources of rice accessions was observed between crosses of any sources (marker repeatability). However, correlation between presence of markers and expression of the traits in rice breeding populations was not significant except for minor portion of traits and markers examined (failure of marker universality). In the present study, various strategies were discussed to develop new markers with universality of breeding application.