鸡卵巢组织SSHcDNA文库构建及差异表达基因筛选与初步鉴定.PDFVIP

吉林农业大学学 2011，05 Journal of Jilin Agricultural University E-mail:jlndxb@ 鸡卵巢组织SSH cDNA文库构建及差异表达基因筛选与初 步鉴定* ** 包艳波，徐日福 ，王志贤，秦 宁，杨雨江，姜建萍 吉林农业大学动物科技学院，长春 130118 摘 要：为分离与鸡产蛋性能密切相关的重要基因或调控因子，利用抑制性消减杂交技术，以高产蛋鸡品种海兰褐商 品代蛋鸡和产蛋量相对较低的地方鸡品种大骨鸡 20 周龄时的卵巢组织为试验材料构建消减 cDNA 文库。文库的插 入片段集中在 500 bp 左右，挑取 720 个克隆进行 PCR 筛选，获得 657 个阳性克隆，经过点杂交筛选后选择了 536 个阳性克隆，从中随机挑取 100 个阳性克隆进行测序。经过同源性比对归并后得到 52 个差异表达基因，除 3 个未知 基因外，其余涉及到细胞繁殖与代谢调控、卵泡生长发育、钙磷和蛋白质营养代谢调节等多个方面。并对 4 个可能 影响鸡产蛋性能的差异表达基因进行了 RT-PCR 半定量检测鉴定。 关键词：鸡；cDNA 文库；卵巢组织；差异表达基因；抑制性消减杂交 中图分类号：S831.2; Q786 文献标识码：A 文章编号：1000-5684(2011) CNKI:22-1100/S1625.001 DOI: 网络出版地址： /kcms/detail/22.1100.s1625.001.html Construction of Suppression Subtractive cDNA Library and Screening and Preliminary Identification of Differentially Expressed Genes in the Ovary of the Different Egg Production Performance in Chicken BAO Yan-bo, XU Ri-fu, WANG Zhi-Xian, QIN Ning, YANG Yu-jiang, JIANG Jian-ping College of Animal Science and Technology, Jilin Agricultural University, Changchun 130118，China Abstract: In order to screen the chief genes and regulatory factors relevant to egg performance in chicken, s uppression subtractive hybridization (SSH) was employed to construct the SSH cDNA library in the ovary from commercial Hyland brown layers of high egg production and Dagu layers of low egg production at 20 weeks of age in the present study. The inserted fragment sizes focused around 500 bp and 720 clones were verified by PCR, in which 657 clones were used to perform dot blotting hybridization. A total of 536 clones were obtained and 100 clones were