• 한국발생생물학회지:발생과생식
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• 제21권3호
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• pp.237-247
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• 2017

Mammalian spermatogenesis occurs in a precise and coordinated manner in the seminiferous tubules. One of the attempts to understand the detailed biological process during mammalian spermatogenesis at the molecular level has been to identify the testis specific genes followed by study of the testicular expression pattern of the genes. From the subtracted cDNA library of rat testis prepared using representational difference analysis (RDA) method, a complimentary DNA clone encoding type III member of a DnaJ family protein, DnaJC18, was cloned (GenBank Accession No. DQ158861). The full-length DnaJC18 cDNA has the longest open reading frame of 357 amino acids. Tissue and developmental Northern blot analysis revealed that the DnaJC18 gene was expressed specifically in testis and began to express from postnatal week 4 testis, respectively. In situ hybridization studies showed that DnaJC18 mRNA was expressed only during the maturation stages of late pachytene, round and elongated spermatids of adult rat testis. Western blot analysis with DnaJC18 antibody revealed that 41.2 kDa DnaJC18 protein was detected only in adult testis. Immunohistochemistry study further confirmed that DnaJC18 protein, was expressed in developing germ cells and the result was in concert with the in situ hybridization result. Confocal microscopy with GFP tagged DnaJC18 protein revealed that it was localized in the cytoplasm of cells. Taken together, these results suggested that testis specific DnaJC18, a member of the type III DnaJ protein family, might play a role during germ cell maturation in adult rat testis.

• 한국어병학회지
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• 제19권3호
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• pp.189-195
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• 2006

A subtracted cDNA library of a marine scuticociliate, Philasterides dicentrarchii, in response to 17β-estradiol exposure was constructed using suppression subtractive hybridization (SSH). As a result of SSH, 275 clones were isolated, and among them, only glutathione peroxidase (GPX) gene was isolated as an antioxidative enzyme responding to 17β-estradiol. The semi-quantitative reverse-transcriptase polymerase chain reaction (RT-PCR) analysis revealed that the transcription of GPX gene of P. dicentrarchii was clearly increased by exposure to 17β-estradiol. The GPX transcription was also clearly increased by exposure to xenoestrogens such as bisphenol A (BPA) and genistein.

• 한국잠사곤충학회지
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• 제50권2호
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• pp.140-144
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• 2012

본 연구는 누에 중장으로부터 면역 관련 유전자를 대량 발굴하고 발현 특성을 분석함으로서 곤충 유래 신기능성 의약품 개발을 위한 유전자 소재를 발굴하고자 하였다. 우선 곤충병원성 섭식에 의한 누에 품종에 따른 중장 면역원으로서 적성 병원성 세균인 X. nematophila 등을 선발하고 누에 천연 면역인자의 발굴을 위해 최적 감염 조건을 설정하였다. 감염된 누에 중장 mRNA를 순수 분리하여 subtraction cDNA 유전자은행 1종씩 제작하고, subtractive differential display hybridization 방법에 의해 누에 중장 면역관련 유전인자를 선발하였다. 선발된 유전자의 정보 분석 결과, 세포 내 다양한 생물학적 기능을 수행하는 것으로 알려진 ribosomal protein L5 mRNA 등 면역 관련 유전자 9종을 선발하였다. 본 연구에서 선발된 누에 천연 면역 관련 인자는 신기능성 의약품 소재로 개발하기 위해서는 기능분석 연구가 지속되어야 할 것으로 사료된다.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제28권1호
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• pp.31-41
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• 2002

Proline-rich proteins (PRPs) are major components of human saliva. In order to know the biological roles of PRPs, we explored the expression pattern and functional protein structures of PRPs by the immunohistochemical and various molecular biological methods. Polyclonal antibody against human gPRP was generated from rabbit by the injection of oral exfoliated cells specially treated by urea and SDS buffer. The PRPs began to be expressed both in the acinar cells and ductal cells from the EIDS (Early Intermediate Developmental Stage) of fetal salivary glands and became intense in the salivary epithelium in the LDS (Late Developmental Stage) and adult salivary glands. The polyclonal antibody against the gPRP showed the cross-reactivity with aPRP and bPRP, these results were relevant to the high homology among subtypes of PRP. However, the simulated protein structures of PRPs showed the characteristic repetitive whorling domains except the N-terminal signal peptide. The whorling domains were also contained the multiple amino acids of glutamine and glycine, which may provide the receptor binding or cross-linking sites of PRPs.

• Asian-Australasian Journal of Animal Sciences
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• 제18권11호
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• pp.1519-1523
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• 2005

By screening a subtracted cDNA library constructed with mRNA obtained from the longissimus dorsi muscles of F1 hybrids Landrace${\times}$Yorkshire and their Yorkshire female parents, we isolated two partial sequences coding for the H3-K4-specific methyltransferase (KIAA1717) and skeletal muscle myosin regulatory light chain (HUMMLC2B) genes. In the present work we investigated two SNPs, one (C1354T) at the 3' untranslated region (UTR) of KIAA1717 and one (A345G) at the SINE (PRE-1) element of HUMMLC2B, in a resource population derived from crossing Chinese Meishan and Large White pig. The selected pigs were genotyped by means of a PCR-RFLP protocol. Significant associations were observed for the KIAA1717 C1354T polymorphic site with thorax-waist backfat thickness (p<0.05), buttock backfat thickness (p<0.05), average backfat thickness (p<0.05), loin eye height (p<0.05), loin eye area (p<0.05), carcass length to 1$^{st}$ spondyle (p<0.01) and carcass length to 1st rib (p<0.01). HUMMLC2B A345G were significantly associated with loin eye width (p<0.05), loin eye area (p<0.05). Further studies are needed to confirm these preliminary results.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.66.2-66
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• 2003

A gain-of-function mutant, SHM-11 obtained through gamma-ray mutagenesis, is resistant to rice blast caused by Magnaporthe grisea while wild type Sanghaehyanghyella is highly susceptible to the same disease. The resistance in the mutant was not race-specific when we tested with four races (KJ-201, KI-1113a, KI-313, KI-409) of M. grisea. To identify genes involved disease resistance in the gain-of-function mutant, genes specifically expressed in the mutant were selected by suppression subtractive hybridization using cDNAS of blast-inoculated mutant and wild type as a tester and a driver, respectively, Random 200 clones from the subtracted library were selected and analyzed by DNA sequencing. The sequenced genes represented three major groups related with disease resistance； genes encoding PR proteins, genes probably for phytoalexin biosynthesis, and genes involved in disease resistance signal transduction. A gene encoding a putative receptor-like protein kinase was identified as highly expressed only in the gain-of-function mutant after blast infection. The role of the putative receptor-like protein kinase gene during blast resistance will be further studied.

• 한국발생생물학회:학술대회논문집
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• 한국발생생물학회 2003년도 전기 한국발생생물학회 제16차 학술대회논문집
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• pp.21-23
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• 2003

Mechanisms regulate the arrest and growth of the resting primordial follicles are very poorly understood. To elucidate genes involved in the early folliculogenesis, we conducted suppression subtractive hybridization using mRNA from day1 and day5 ovaries and selected weel for further analysis, since it was most frequent gene in the day1-subtracted cDNA library (1). Expression of weel and correlated components of the cell cycle machinery, such as cdc2, cyclin B1, cdc25C, and phosphorylated cdc2 was evaluated by immunohistochemistry. In primordial follicles, expression of weel, cdcw, and cyclin B1 was cytoplasmic in oocytes, but phosphorylated cdc2 was weakly expressed in oocytes. While cdc25C expression was in ovarian somatic and in some theca cells. None of components was expressed in the pre-granulosa cells of the primordial follicles, while weel weakly, and cdc2 and cyclin B1 was strongly expressed in the granulosa cells of the growing follicles. Results from the present study suggest that 1) the mejotic arrest of the oocytes may not due to of cell cycle machinery, and 2) the weel may arrest meiosis by sequestering cdc2 and cyclin B1 in the cytoplasm by protein-protein interactions and/or by inhibitory phosphorylation.

• Clinical and Experimental Reproductive Medicine
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• 제33권4호
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• pp.253-263
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• 2006

목 적: 본 연구에서는 EST Clustering 방법을 이용하여 이전의 연구에서 발굴한 B357 EST의 염기서열을 포함하는 유전자를 동정하고, 이 유전자의 발현을 생쥐의 난소와 정소를 포함한 여러 가지 조직들에서 살펴보고자 하였다. 연구방법: EST Clustering으로 얻어진 전체 염기서열을 5-day-ovary-specific gene-1 (5DOS1)이라고 명명하여 GenBank에 등록하였으며 (AY751521), northern blotting, real-time RT-PCR, in situ hybridization, western blotting, immunohistochemistry 등의 방법을 이용하여 생쥐 난소와 고환의 발달단계에 따라 그 발현양상을 관찰하였다. 결 과: 5DOS1의 전사체는 성장한 정소, 뇌, 근육에서 높게 발현하였으며, 난소의 경우에서는 원시난포시기부터 모든 난자에서 발현하였으며 특히 생후 5일째 높게 발현하였고 그 이후로는 점차 감소하였다. 반면 정소의 경우는 발달과 함께 계속 증가하였으며, 정모세포를 제외한 모든 발달단계별 정자에서 발현함을 관찰하였다. 결 론: 본 연구결과는 5DOS1에 대한 발견과 동정에 대한 첫 보고로써, 유전자 및 단백질이 생쥐의 난소 및 정소의 생식세포에서 발현하는 것을 관찰하였다. 앞으로 생식세포 발생 및 분화에 관련된 5DOS1의 기능에 대한 심층 연구가 더 필요하다고 사료된다.

• 한국발생생물학회지:발생과생식
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• 제4권1호
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• pp.67-77
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• 2000

심해에서 서식하며 특이한 조직형태를 갖고 있는 꼼치조직에서 cDNA를 제작하여 클로닝을 통해서, 꼼치에서 특이하게 발현하는 유전인자를 검색하였다. 그 결과 62례의 클론이 알려진 유전자에 동질성이 있었는데 이들은 thioesterase가 9례, myosin이 8례, creatine kinase가 7례, skeletal alpha-actin이 6례, parvalbumin b와 ribosomal protein이 각각 5례, type I collagen과 muscle troponin이 각각 3례, dopamine receptor, histatin, 그리고 heat shock protein이 각각 2례, cystatin, lectin, statherin, secretory carrier membrane protein, keratin type I, desmin, chloroplast, muscle tropomyosin, reticulum calcium ATPase, ribonucleoprotein이 각각 1례로 나타났다. 나머지 클론은 동질성이 낮거나 비반복성으로 나타났으며, 이 중 in situ hybridization으로 조직에서 특이하게 발현되는 5종류의 클론을 선택하여 분석하였으며, C 말단 단백질 구조와 특색(motif)을 분석하였다. 5종류의 클론에서 C9O-77은 약 5000bp 크기로 상피조직, 점액조직, 섬유조직 그리고 교원질 조직에서 강한 양성반응을 보이는 세포의 기질단백질로 예상되었다. C9O-116은 약 1500bp 크기로 섬유조직, 상피조직 그리고 점액조직에서 약한 양성반응을 보였고, 근육 다발 주변 부위 세포에서 매우 강한 양성반응을 보이는 막투과성 단백질로 예상되었다. C9O-130은 약 1200bp 크기로 상피조직, 근육조직 그리고 점액조직에서 양성반응을 나타냈으며, 특히 상피조직에서 강한 양성반응을 나타내는 세포내 단백질로 예상되었다. C9O-161은 약 2000bp 크기로 상피조직 과 근육조직 그리고 점액성 섬유조직에서 약한 양성반응을 보였고, 상피세포에서 강한 양성반응을 보였으며, 근육다발을 둘러싸는 섬유성 세포에서도 강한 양성반응을 보이는 세포외 기질단백질로 추측되었다. C9O-171은 약 1000bp크기로 상피조직, 근육조직 그리고 섬유기질 조직에 널리 강한 양성반응을 나타냈으며 교원띠조직에서도 양성반응을 보이는 전사인자로 추측되었다.