籽鹅卵巢组织全长cDNA文库的构建及部分克隆序列分析.pdfVIP

农业生物技术学报，2010 年，第18 卷，第2 期，第389-393 页 Journal of Agricultural Biotechnology, 2010, Vol.18, No.2, 389-393 研究资源 A Resource 籽鹅卵巢组织全长cDNA 文库的构建及部分克隆序列分析 1 2 3 1* 董重阳 康波 贾晓剑 杨焕民 1 黑龙江八一农垦大学动物科技学院, 大庆, 163319; 2 四川农业大学动物科技学院, 雅安, 625014; 3 黑龙江省青年农场畜牧科, 哈尔滨, 150050 * 通讯作者, yanghuanmin@ 摘 要 为克隆鹅产蛋性能相关基因，以籽鹅( )卵巢组织为实验材料，以 TriplEx2 为载体，用 Anser anser λ SMART (switching mechanism at 5 end of RNA transcript)方法构建了籽鹅卵巢组织的全长cDNA 文库。结果 6 表明，该文库初始滴度为3.1×10 pfu/mL ，重组率为94%，插入片段长度集中在0.5~2 kb。随机挑取120 个克 隆测序，测序成功110 个，分别对应于81 个cDNA 片段，与BLASTn 进行序列比对，其中有70 个为已公布 的EST ，其中大部分与原鸡或其它物种具有较高的同源性，而11 个cDNA 与其它物种之间同源性低，可能 是未知功能基因。 关键词 籽鹅, 卵巢, cDNA 文库 Construction of the Full-length cDNA Library and Analysis in Part of ESTs in Zi Goose Ovary Dong Chongyang 1 Kang Bo 2 Jia Xiaojian 3 Yang Huanmin 1* 1 College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University, Daqing, 163319; 2 College of Animal Science and Veterinary Medicine, Sichuan Agricultural University, Yaan, 625014; 3. Department of Animal, Heilongjiang Youth Farm, Harbin, 150050 * Corresponding author, yanghuanmin@ DOI: 10.3969/j.issn.1674-7968.2010.02.030 Abstract In order to clone the full-length of gene related to egg laying performance, the experiment was con- ducted to constructed the full-length cDNA library of the ovary in the Zi geese (Anser anser) by SMART (switching mechanism at 5 end of RNA transcript) by λTriplEx2 as vector. The results showed that the titration of the cDNA 6 library was 3.1×10 pfu/mL, the rate of recombination was 94%, and the fragment length of inserted DNA ranged mainly from 0.5 kb to 2 kb. One hundred and tenty clones were chosen at random for sequencing from the cDNA library, and 110 clones were sequenced successfully, and after BLASTn ana