In the present study, the plantlets in vitro of Paeonia suffruticosa ‘Wu Long Peng Sheng’ were used as laboratory materials. The proteome during adventitious root induction process was investigated to sift the related proteins by two-dimensional electrophoresis and mass spectrometry. The results indicated that the protein spots were concentrated in the acidity gel region (pH 4 - 7) and the spots number had a dynamic change ranged from 373 to 462 at the process of root induction (0 – 7 d). 8 spots significantly changed were analyzed with a mass spectrometer and identified using associated software and databases. The peptide information of the 8 spots was similar to the ATP synthase β-subunit of P. suffruticosa (Spots 1 - 4 and 8), P. tenuifolia (Spots 5), P. californica (Spot 6) and P. brownie ( Spots 7) r espectiv ely. T he expression levels of protein spots 1, 4, 5, 6 and 7 was dramatically downregulated, and that of protein spots 2 and 3 had a slightly opposite tendency on the 3rd day. The obviously decreased period is particularly interesting as it was consistent with the induction period of adventitious root primordial of tree peony plantlet in vitro. The ATP synthase β-subunit could be consumed for assembling the ATP synthase in order to supply energy to the rooting process. Therefore, we speculated that the ATP synthase β-subunit was involved in adventitious root initiation of tree peony plantlets in vitro and we expect that further studies should be carried out in order to export its action mechanism.

Nuts are one of the most common sources of allergies in individuals of all ages. In order for a particular protein to render an allergic reaction, it must resist proteolytic digestion by intestinal enzymes. In this study, three well-known allergenic nuts, almonds, cashew nuts, and peanuts, were used as samples, and enzyme digestion with Bacillus protease and porcine pepsin was tested. A proteomic approach using two-dimensional gel electrophoresis and an MS/MS analysis was applied to visualize and identify the proteins that were resistant to enzyme digestion. Among the 150 protein spots tested, 42 proteins were assigned functions. Due to the lack of genomic databases, 41% of the identified proteins were grouped as hypothetical. However, 12% of them were well-known allergens, including AraH. The remainder were grouped as storage, enzymes, and binding proteins.

This study was to evaluate the protein profile of seminal plasma using 2-DE in Hanwoo. Seminal plasma was harvested from five mature Hanwoo, and seminal plasma protein was extracted by M-PER Mammalian Protein Extraction Reagent. Proteins were refined by clean-up kit and quantified by Bradford method until total protein was . Immobilized pH gradient (IPG) strip was used 18 cm and 3~11 NL. SDS-PAGE was used 12% acrylamide gel. Each gels were visualized by comassie brilliant blue and silver staining. These spots were analyzed by MALDI-TOF MS and searched on NCBInr. The result, 20 proteins of 36 protein spots were searched through peptide sequencing on the NCBInr. 8 proteins profiled by 2-DE were proved through previous bovine studies and the name of each protein was albumin, nucleobindin, clusterin, TIMP-2, spermadhesin Z13, spermadhesin-1 and BSP proteins (BSP 30 kDa and BSP A1/A2). 12 new proteins were ATP synthase, protein MAK16 homolog, Transmembrane protein 214, E3 ubiquitin-protein ligase BRE1A, dual serine/threonine and tyrosine protein kinase, tissue factor pathway inhibitor 2, alpha-actinin-4, RUN domain-containing protein 3B, catenin alpha-1, protein-glutamine gamma-glutamyltransferase 2, plakophilin-1 and inter-alpha-trypsin inhibitor heavy chain H1 has not been previously described in the bovine seminal plasma study. These proteins may be contribute to define the type of proteins affecting fertility of male and improve the fertilizing ability of semen in Hanwoo.

When fully grown oocytes are removed from their follicles, they can resume meiosis and mature spontaneously under in vitro conditions. However, nuclear maturation under in vitro condition is not accompanied by complete cytoplasmic maturation, which is essential for successful fertilization and the initiation of zygotic development. This study analyzed change of proteins in follicular fluids during the porcine follicular development. Follicular fluids were collected from follicles of diameter 1～2 mm, 2～6 mm and 6～10 mm in ovary of slaughtered pigs. Total proteins were extracted from follicular fluids by M-PER Mammalian Protein Extraction Reagent. We confirmed totally 27 same spots, 1 spot from follicle fluid of 2～6 mm follicle and 5 spots from follicle fluid of 6～10 mm in diameter were analyzed by MALDI mass spectrometry and searched on NCBInr. In results, spot No. 28 from 2～6 mm follicle was Ig lambda chain C region, and spot No.32 and 33 from 6～10 mm was Apolipoprotein A-(APOA4). Spot No.29 and 31 were failed to analyze. These results indicate that the porcine oocyte during in vitro maturation depend on specific different expressed proteins may play an important roles in the sequence of molecular events in porcine oocyte maturation and follicular development.

Changing Proteins in Granulosa Cells during Follicular Development in Pig In-Soon Chae1, Dong-Min Jang3, Hee-Tae Cheong2, Boo-Keun Yang1 and Choon-Keun Park1,† 1College of Animal Life Sciences, 2School of Veterinary Medicine, Kangwon National University, Chuncheon 200-701, Korea 3CHA Stem Cell Institute, CHA Hospital, Seoul 463-712, Korea ABSTRACT This study analyzed change of proteins in granulosa cells during the porcine follicuar development by proteomics techniques. Granulosa cells of the follicles, of which the diameter is 2～4 mm and 6～10 mm, were collected from ovary of slaughtered pig that each follicle of diameter 1～4 mm and 6～10 mm. We extracted glanulosa cell proteins by M-PER Mammalian Protein Extraction Reagent. Proteins were refined by clean-up kit and quantified by Bradford method until total protein was 200 μl. Immobilized pH gradient(IPG) strip used 18 cm, 3～10 NL. SDS-PAGE used 10% acrylamide gel. After silver staining, Melanie 7 and naked eye test were used for spot analyzation. Increasing proteins in glanulosa cell of 6～10 mm follicle were 7 spots. This spots were analyzed by MALDI-TOF MS and searched on NCBInr. In results, 7 spots were similar to zinc/ling finger protein 3 precursor (RING finger protein 203), angiomotin, heat shock 60 kDa protein 1 (chaperonin) isoform 1 (HSP60), similar to transducin-like enhancer protein 1 (TLE 1), SH3 and PX domains 2A (SH3PXD2A). Those proteins were related with transfer between cells. Increase of proteins has an effect on follicular development.

The early diagnosis of bovine pregnancy is an essential component of successful reproductive planning on farms, because lack of bovine pregnancy over the long term results in reproductive failure and low milk yield‐the latter of which is a special concern on dairy farms. This study was designed to identify early pregnancy‐specific whey proteins in bovine, by comparing milk samples collected from cattle during pregnancy (Days 30 and 50) and from non‐pregnant cattle. In this study, differentially expressed proteins in five pregnant and five non‐pregnant Holstein dairy cattle were investigated and compared, using proteomics analysis. The first dimension was applied to a pH 3.0～10.0 strip, by loading a 2‐mg milk protein sample. After the second‐dimension separation was performed, the gels were stained with colloidal Coomassie brilliant blue. The stained gels were scanned and the images were analyzed, to detect variations in protein spots between non‐pregnant and pregnant cattle milk protein spots, using ImageMaster; this was followed by analysis with MALDI TOF‐MS. Analysis of the 2‐DE gel image resulted in a total of approximately 500～600 protein spots, of which 12 spots were differentially expressed, six spots were up‐regulated, and four spots were downregulated; two spots were identified as pregnancy‐specific proteins. These proteins were identified as lactoferrin, NADH dehydrogenase subunit 2, albumin, serum albumin precursor and transferrin. Our results via 2‐D PAGE analysis revealed composite profiles of several milk proteins related to early bovine pregnancy, implying the possible use of these milk proteins in the early detection of bovine pregnancy.

To investigate the antifungal activity related protein in pesticidal bacteria, a bacterial strain LTD was isolated from soil collected at Gimje in Jeonbuk province, Korea, and identified as Bacillus subtilis LTD based on a API50 CHB kit and 168 rDNA seque

The proteins from functional rice cultivars (Nogwonchalbyeo, Giant embryonic, Arhyangchalbyeo, and Goamibyeo) and general white rice were extracted and separated using two-dimensional (2D) gel electrophoresis. A wide variation in the molecular weight (MW) and pH range of the expressed proteins in rice samples were observed. The green-kerneled rice (Nogwonchalbyeo) exhibited proteins with MW of 9-57 kDa and appeared at a pH range of 4-7. The Giant embryonic contained proteins with MW of 31-63 kDa and a pH range of 5-6. The aromatic glutinous rice (Arhyangchalbyeo) showed proteins with MW of 24-28 and pH of 5.8-6.8. The high-amylose rice (Goamibyeo) exhibited proteins with MW of 3-63 and pH of 5.2-5.6. The identified proteins uniquely found and highly expressed in each cultivar may have a significant role on rice functionality. The results illustrate that the 2D gel electrophoresis is a valuable method in the determination of the protein expression profiles in functional rice grains and may be useful in the identification of specific marker proteins associated with the functional property of rice.

이차원 전기영동 분석을 이용하여 국내 밀 32 품종의 HMW-GS 단백질 발현의 정성 및 정량적인 분석을 통해 품 HMW-GS 발현 정도를 평가하여 국내 밀 품종 육성의 초 자료로 활용하고자 수행하였다. 평균 HMW-GS 스팟 수는 11.78개였으며, Glu-A1 1.31개, Glu-B1 5.53개, 그리고 Glu-D1에서 4.94 개였다. Glu-B1과 Glu-D1에서는 subunit에 따른 단백질 스팟 수가 차이가 없기 때문에, Glu-A1에서는 1과 2* subunit을 지닌 품종이 null allele 품종에 비하여 단백질 스팟 수가 많았다. 단백질 스팟 수는 조경밀이 18개로가장 많았으며, 다홍밀은 7개로 제일 적었다. 단백질의 상대적인 발현량을 조사한 결과 평균 0.44로 대비 품종인 Chinese Spring에 비하여(1.0) 낮았고, 고분밀이 1.11로 가장 높았으며, 은파밀이 0.24로 가장 낮았다. 단백질 스팟수와 발현량을 이용한 유연관계 분석 결과, 국내 밀 품종을 6개 그룹으로 분류할 수 있었다.

To evaluate expression level of HMW-GS protein qualitatively and quantitatively, we separated glutenin fractions and conducted two-dimensional electrophoresis (2DE) in 32 cultivars of Korean wheat for the use of as the basis of wheat breeding. The average spot number of HMW-GS in all Korean wheat cultivars was 11.78 which included 1.31, 5.53 and 4.94 to Glu-A1, Glu-B1 and Glu-D1 loci, respectively. Cultivars harboring 1, 2* subunits had many spots more than ones harboring null allele in Glu-A1 loci because there is no difference of spots between Glu-B1 and Glu-D1 loci. In total spot number of HMW-GS, the highest one was Jokyung as 18 and Dahong the lowest as 7. When the Korean wheat cultivars were compared with the Chinese spring in the average relative expression level, Korean one’s were lower as 0.44. Especially, Gobun was the highest as 1.11 and Eunpa was the lowest as 0.24. Also we investigated phylogenetic relationship based on both frequency of HMW-GS spots and quantification value of each spot to all HMW-GS spots. As a result, Korean the varieties of Korean wheat could be classified into six groups.

LMW-GSs represent approximately 1/3 of the total wheat gluten fraction, which have not been widely studied, even though they are important in the context of wheat end-use quality. In this study, we report on the qualitative and quantitative analysis of LMW-GS in korean wheat cultivars by 2DE in 32 cultivars of Korean wheat for the use of the basis of wheat breeding. We firstly identified spots corresponding each of Glu-3 alleles. The 2DE results for each cultivar will be used as reference map or protein marker discriminating wheat cultivars, wheat and rice, imported and Korean flour. Unexpectedly, five LMW-GS spots were found to be expressed at a common position in hexaploid wheat cultivars, and these spots might play something in glutenin biosynthesis. Total spot numbers were expressed variously between 20 and 10, and average spot number was shown 17.12. The average number of spots in Glu-A3, Glu-B3 and Glu-D3 were 3.0, 4.56 and 2.96 respectively. When the Korean wheat cultivars were compared with the Chinese spring (1.0) in the average relative expression level, Korean one’s were lower as 0.67. Especially, Gobun was the highest as 1.32 and Baekjoong was the lowest as 0.24. Also we investigated phylogenetic relationship based on frequency of HMW-GS spots and quantification value of each spot to all LMW-GS spots. As a result, the varieties of Korean wheat could be classified into five groups.