草业学报2012，Vol.21Issue(1)：296-301,6.
象草4CL基因片段的克隆及RNAi表达载体构建
Cloning of a 4CL gene fragment and construction of a RNAi expression vector of Pennisetum purpureum
摘要
Abstract
Pennisetum purpureum is a perennial resource plant and widely cultivated in tropical and sub-tropical regions. A pair of specific primers was designed to assess the effect of down-regulated expression of the 4-cou-marate: Co A ligase (4CL) gene on lignin composition of P. purpureum cv. Huanan, using the 4CL gene sequence of P. purpureum cv. Mott (accession No: GU997597. 1). An interference fragment of 374 bp was obtained by PCR amplification with the above primers and it was cloned into the pCR /GW/TOPO vector by TOPO cloning to obtain pENTER-4CL. The corresponding fragment on destination vector pCB2004B was then replaced by the interference fragment of pENTR-4CL by LR reaction to form the RNAi (RNA interference) expression vector pCB2004B-4CL. The Agrobacterium-mediated transformation of the expression vector was carried out using the freeze-thaw method, and the expected fragments appeared in the PCR products using bacterial liquid as a template. This study was a basis for research on the 4CL function of P. purpureum by RNAi technology.关键词
象草/Gateway技术/4CL基因/RNAi/载体构建Key words
Pennisetum purpureum/ Gateway technique/ 4CL gene/ RNA interference/ vector construction分类
农业科技引用本文复制引用
霍松,陈慧,朱琼华,解新明..象草4CL基因片段的克隆及RNAi表达载体构建[J].草业学报,2012,21(1):296-301,6.基金项目
国家自然科学基金项目(30972138和30671489)资助. （30972138和30671489）
