Abstract:Objective To clone promoter fragment of Toxoplasma gondii LDH2 gene and to evaluate the promoting activity. Methods A series of fragments of 5′UTR of Toxoplasma gondii LDH2 gene were amplified by PCR, then inserted into pTX3-Basic via Kpn I and Xhol I to construct luciferase report plasmids. Forty eight hours after tachyzoites being co-transfected by luciferase report plasmids and reference plasmid pRL-Tg,dual-luciferase assays were carried out to determinate activities of the fragments to initiate luciferase transcription. Results The sequence of the 5′UTR of Toxoplasma gondii LDH2 gene fragments proved to be correct by sequencing and a series of luciferase report plasmids were verified by PCR, restrictive endonuclease digest and sequencing. The luciferase activities of all the luciferase report 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, 1.2 to 1.5 folds to the positive control pTX3-Control except the plasmids pTX3-293 and pTX3-193. Conclusion The promoter region of Toxoplasma gondii LDH2 was located and cloned successfully. Our results provide evidence to investigate the mechanism of regulation of the LDH2 gene.
收稿日期: 2012-08-15
出版日期: 2013-06-19
基金资助:国家自然科学基金;福建省自然科学基金;福建省卫生厅青年基金
通讯作者:
侯香华
E-mail: xianghuahou@yahoo.com
引用本文:
张加勤 宋秀宇 侯香华. 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.
Jiaqin Zhang. CLONING OF TOXOPLASMA GONDII LDH2 GENE PROMOTER AND EVALUATION OF ITS PROMOTING ACTIVITY. Chinese Journal of Zoonoses, 2013, 29(4): 349-353.