中国人兽共患病学报2013，Vol.29Issue(4)：349-353,365,6.DOI:10.3969/cjz.j.issn.1002-2694.2013.04.007
弓形虫乳酸脱氢酶LDH2基因启动子的克隆及鉴定
Cloning of Toxoplasma gondii LDH2 gene promoter and evaluation of its promoting activity
摘要
Abstract
To clone promoter fragment of Toxoplasma gondii LDH2 gene and to evaluate the promoting activity, a series of fragments of 5 UTR of Toxoplasma gondii LDH2 gene were amplified by PCR, and then inserted into pTX3-basic via Kpn I and Xhol I to construct luciferase reporter plasmids. Forty eight hours after tachyzoites being co-transfected by luciferase reporter plasmids and reference plasmid pRL-Tg, dual-lucif erase assays were carried out to determinate activities of the fragments to initiate luciferase transcription. Results showed that the sequence of the 5 UTR of Toxoplasma gondii LDH2 gene fragments was proved to be correct by sequencing, and a series of luciferase reporter plasmids were verified by PCR, restrictive endonuclease digest, and sequencing. The luciferase activities of all the luciferase reporter plasmids with 5 UTR of Toxoplasma gondii LDH2 in tachyzoites were similar with the negative control pTX3-basic. While in bradyzoites, all the plasmids yielded 40 to 50 folds of luciferase activity to the negative control, and 1. 2 to 1. 5 folds to the positive control of pTX3-basic, excepting for the plasmids pTX3-293 and pTX3-193. In conclusion, the promoter region of Toxoplasma gondii LDH2 was located and cloned successfully. Our results provide evidence for investigation of the mechanism on regulation of the LDH2 gene.关键词
弓形虫/乳酸脱氢酶/启动子/荧光素酶Key words
Toxoplasma gondii / LDH2 / promoter/ luciferase分类
医药卫生引用本文复制引用
张加勤,侯香华,宋秀宇..弓形虫乳酸脱氢酶LDH2基因启动子的克隆及鉴定[J].中国人兽共患病学报,2013,29(4):349-353,365,6.基金项目
国家自然科学基金(No.81000762) （No.81000762）
福建省自然科学基金(No.2010D018) （No.2010D018）
福建省卫生厅青年基金(No.2010-2-90) （No.2010-2-90）
