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OALib Journal期刊
ISSN: 2333-9721
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锯缘青蟹性腺发育与性别分化相关组织混合线性化cdna文库构建及est初步分析
Keywords: 锯缘青蟹scyllaserrata,线性化cdna文库,性腺发育,性别分化est
Abstract:
？分别提取锯缘青蟹scyllaserrata卵巢、精巢、眼柄、促雄腺的总rna，等量混合后经oligotex试剂分离mrna，利用smart(switchmechanismsatthe5’endofrnatranscript)技术及dsn(duplex-specificnuclease)的特性构建线性化cdna质粒文库。经检测文库约含有5.3×107重组子，扩增后的文库含有1011以上重组子。文库的插入cdna长度为0.5-2.5kb，重组率大于98%。从构建的文库中随机挑取5202个克隆进行大规模est测序，获得3346条ests，其中长度大于100bp的高质量ests2697条。经拼接与软件分析，2697ests代表了2522个unigenes，包括2355个singlets和167个contigs，冗余率6.49%。est序列经生物信息学分析(blastx,e<10-6)，发现了sry-likeproteinc、sox14protein、sox4b、sex-determiningproteinfem-1、vitellogenin、vitellogeninreceptor、cyclina、cathepsinc和cyclin-dependentkinase2等与性别分化和性腺发育相关的基因，以及热休克蛋白家族、泛素系统、抗氧化防御体系和抗病相关的功能基因。该文库具有较高的质量和利用价值，可为更大规模est分析及采用基因芯片技术开展锯缘青蟹性腺发育及性别分化相关功能基因的研究奠定基础。
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