Recruitment of primordial follicles(PMF) is crucial for female fertility. however, factors and mechanisms that regulate this process is poorly understood. The present study was conducted to obtain an inclusive view of the gene expression and to identify novel factors and their pathways of regulating PMF arrest and/or growth initiation. Ovaries from one-day neonatal(consists of oocyte and PMF) and five-day old(consists of PMF and primary follicles, PRIF) mice were collected, either total RNA or mRNA was isolated, and suppression subtractive hybridization(SSH) was used to isolate and clone genes that differentially expressed in day 1 and day 5 ovaries. Confirmation that some of these genes are differentially expressed in PMF and/or in PRIF was accomplished by using laser captured microdissection(LCM), RT-PCR. in situ hybridization(ISH) and/or immunohistochemistry(IHC). In toto, 357 clones were sequenced and analyzed by BLAST and RIKEN program. Sequences of 330 clones significantly matched database entries while 27 clones were novel. Forty-two and 47 different genes were identified as differentially expressed in day 1 and day 5 ovaries, respectively, while 7 genes were expressed in both stages of ovaries. Day 5-subtracted library included several genes known as markers far growing follicles, such as ZP2, MATER, and fetuin. Among the genes with assigned functions, 23.8% was associated with cell cycle/apoptosis regulation, 7.1% with cellular structure, 11.9% with metabolism, 26.2% with signal transduction, and 31.0% with gene/protein expression in day 1; while 10.6%, 17.0%, 23.5%, 25.5%, and 23.4% in day 5, respectively. Genes such as GDF-8, Lats2, Septin2, and Weel were the highly expressed genes in PMF, while HSP84, Laminin2, MATER, MTi7, PTP, and Wrn were highly expressed genes in PRIF. We have successfully discovered list of genes expressed in day 1 and day 5 ovaries and confirmed that some of them are differentially expressed in PMF and/or PRIF. Gene expression profile from the present study would provide insight for the future study on the mechanism(s) involved in primordial-primary follicular transition. This work was Supported by Korean Health 21 RND Project, Ministry of Health and Welfare, Korea (01-PJ10-PG6-01GN13-0002).

본 연구는 V. nashicola에 의해 발생하는 배 검은별무늬병 에 저항성을 가지는 유전자를 선발하고자 고도 저항성 ‘93-3-98’과 고도 감수성 ‘스위트스킨’에 병 접종후 24시간, 48시간후 엽을 채취하여 suppression subtractive hybridization 을 수행하였다. ESTs 분석결과 ‘93-3-98’에서만 발현된 유전 자는 24H(tester, 24시간 ’93-3-98; driver, 24시간 ‘스위트스킨’)과 48H(tester, 48시간 ’93-3-98; driver, 48시간 ‘스위트 스킨’)에서 각각 9개, 14개였으며 방어 또는 스트레스에 관련 된 유전자의 비율은 각각 40%와 42%였다. 식물의 방어반응에 관련된 PR protein family 유전자로써는 24H에서 pathogenesisrelated protein 1a, major allergen Pyr c 1과 allergen mal d 1, 48H에서는 major allergen Mal d 1.03B 유전자가 특이 적으로 발현되었고 다수의 방어반응에 관련된 유전자들도 확 인되었다. Major allergen Pyr c 1, F-Box/kelch-repeat protein, flavoprotein wrbA, hypothetical protein POPTRDRAFT_ 783792은 고도저항성 ‘Bartlett’과 ‘93-3-98’에서만 높게 발 현 되었으며 중도저항성 ‘감천배’와 감수성 ‘원황’, 고도 감수 성 ‘신고’와 ‘스위트스킨’에서는 낮게 발현되어 검은별무늬병 저항성 연관 유전자로 확인되었다.