凡纳滨对虾兰尼定受体基因亚克隆文库的构建及部分序列的测定

凡纳滨对虾兰尼定受体基因亚克隆文库的构建及部分序列的测定 Construction of Subclone Library of Litopenaeus vannamei Ryanodine Receptor and Determination of Its Partial Sequence 陈晓汉 CHEN Xiaohan 1 first-author 马宁 MA Ning 1 曾地刚 ZENG Digang 1 广西水产研究所 广西水产研究所 Fisheries Research Institute of Guangxi Fisheries Research Institute of Guangxi 将一个含有凡纳滨对虾兰尼定受体基因的fosmid克隆用DNA剪切仪进行片段化处理,回收3～5kb之间的DNA片段连接到pUC118HincⅡ/BAP载体上,构建了适合于鸟枪法测序的亚克隆文库.经鉴定,该文库滴度为5.0×104 cfu.mL-1,重组率达到90%,插入片段大小在3～5kb范围内.随机挑取100个阳性克隆进行两端测序,通过拼接得到一条兰尼定受体基因部分片段,该片段长2 550bp.通过比对分析,发现该基因片段推导的氨基酸序列与果蝇兰尼定受体基因有较高的相似性. A fosmid clone containing the Litopenaeus vannamei ryanodine receptor（RyR） gene was cut by DNA shearing instrument,and the 3-5 kb fragments were collected and ligated with pUC118 HincⅡ/BAP to construct a subclone library suitable for shotgun sequencing.The titer of the library was 5.0×104 cfu·mL-1,and the recombination rate of it was 90%.The inserted fragments were in the range of 3-5 kb.One hundred positive subclones were isolated and double end-sequenced and assembled.Then a RyR gene fragment,2 550 base pairs in size,was obtained.Blast analysis showed that the deduced amino acid sequence of this fragment was highly similar to the amino-terminal sequences of Drosophila melanogaster ryanodine receptor. 凡纳滨对虾 亚克隆文库 兰尼定受体基因 Litopenaeus vannamei subclone library ryanodine receptor gene 10.14188/j.1671-8836.2011.01.015 S917.4 国家自然科学基金（30860214）;国家岗位科学家经费（nycytx-46）;广西自然科学基金（0991106）;广西科技攻关项目（0992025-3）资助项目 2011-01-01 2021-04-01 1 57