深圳大学学报（理工版）2012，Vol.29Issue(6)：534-540,7.DOI:10.3724/SP.J.1249.2012.06534
一种莱茵衣藻启动子功能检测系统的构建
Construction of Chlamydomonas reinhardtii system for analyzing the function of algal promoter
摘要
Abstract
Chlamydomonas reinhardtii was employed as a bio-system for analyzing the function of algal promoter in this study. The C. reinhardtii T-vector based on pSP124 plasmid was constructed using 450 bp sequences of β-carotene ketolase gene ( bktl) promoter as insert fragment. Two Eaml 105 Ⅰ restriction sites were introduced into this T-vector. 1 986 bp sequences of bktl promoter was obtained by PCR and cloned into the C. reinhardtii T-vector directly. By "glass-bead method" , transformants of Tran and Tran Ble were generated from TAP containing 10 (μg/mL Zeomycin. However, none of Tran B-2 were observed due to some negative regulatory elements in 1 986 hp-bkt 1 promoter. Results of PCR confirmed that ble was integrated into the genome DNA of C. reinhardtii . The 450 bp sequences of bktl promoter were able to express BLE in transgenic algae, which verified that it owned promoter ability in C. reinhardtii. These results indicate that the C. reinhardtii and pB-0. 45T system for analyzing promoter function is workable. It is a new way for studying algal promoters.关键词
基因工程/T载体/莱茵衣藻/启动子功能/检测系统/珠磨法/克隆Key words
genetic engineering/ T-vector/ Chlamydomonas reinhardtii/ promoter function/ analyzing system/ glass-bead method/ cloning分类
生物科学引用本文复制引用
王潮岗,黄惠珠,孙海珊,胡章立,雷安平..一种莱茵衣藻启动子功能检测系统的构建[J].深圳大学学报（理工版）,2012,29(6):534-540,7.基金项目
国家自然科学基金资助项目(31000162,31070323) （31000162,31070323）
