桑树叶绿体基因组DNA的提取及部分序列分析  

A Rapid cpDNA Isolation of Mulberry, Moris L. and Analysis of Part Sequence

作　　者：赵卫国[1] 赵巧玲[1] 张志芳[1] 肖庆利[1] 潘一乐[1] 何家禄[1] 

机构地区：[1]农业部家蚕生物技术重点开放实验室,中国农业科学院蚕业研究所,镇江212018

出　　处：《蚕业科学》2001年第4期303-305,共3页ACTA SERICOLOGICA SINICA

摘　　要：改进Milligan法 ,提取了桑树叶绿体基因组DNA(cpDNA)。利用特异性引物 ,从cpDNA基因组扩增出tRNL tRNF基因。再用随机引物对同一种桑基因组DNA及cpDNA进行RAPD分析 ,表明提取的DNA是具有相当纯度的cpDNA。进一步用XbaⅠ和HindⅢ进行了cpDNA的酶切和克隆。通过酶切鉴定 ,已克隆到 5个片段 ,对其中 1个片段的 70 0bp序列进行了分析 ,推测克隆的片段可能为trnK基因的内含子。According to modified Milligan method, the chloroplast genomic DNA of mulberry was prepared. The result of amplification with special primer of tRNL and tRNF and random primer indicated that the extracted DNA was chloroplast genomic DNA of mulberry. In addition, the cpDNA of mulberry was digested by Xba Ⅰ and Hin dⅢ restriction endonucleases and 5 restriction fragment were cloned, of which the 700bp fragment was sequenced. Sequence analysis showed that cloned 700bp fragment was probably the intron of trn K gene of mulberry chloroplast.

关 键 词：桑树 叶绿体 基因组DNA 序列分析 提取 基因克隆 

分 类 号：S888.2[农业科学—特种经济动物饲养]