Effects of Tetracycline Operator Element Insertion on Plasmodium falciparum Glycophorin Binding Protein 130 Gene Promoter Activity

Abstract

Abstract: Objective To construct tetracycline operator (TetO) modified glycophorin binding protein 130 gene (GBP130) promoter of Plasmodium falciparum and investigate the position effect of insertion of TetO on the promoter activity. Methods Cloning of 7-copy of TetO (7cot) sequence into 4 points relative to transcriptional initiation site of GBP130 promoter in pGBPCATΔ2 plasmid (2 upstream and 2 downstream), respectively, produced 4 derivative plasmids, pG/7T(-5), pG/7T(-2), pG/7T(+2) and pG/7T(+5). After transient transfection, the expression level of reporter gene CAT in both pGBPCATΔ2 and its derivative plasmids was detected and analysed by CAT ELISA. Results Identification by enzyme restrictions and PCR amplifications, as well as DNA sequencing confirmed that the plasmids were successfully constructed. Transfection of these plasmids and CAT detection suggested that insertion of 7cot into each point of GBP130 promoter enhanced the promoter activity, and downstream location of 7cot in the promoter showed higher promoter activity than those located upstream, with the strongest effect in Ins 4 (point +5). Conclusion Plasmid pG/7T(+5), in which 7cot was inserted into +5 point of GBP130 promoter, can be chosen as the responsive plasmid in establishing tetracycline-controlled transgenic expression system of malarial parasites.

Key words: Plasmodium falciparum, glycophorin, carrier proteins, tetracycline, transfection

WANGXian-feng;XUECai-fang;LIUZhong-xiang;LIXun;LIShu-mei;LEIJun-chuan;MIAOJun. Effects of Tetracycline Operator Element Insertion on Plasmodium falciparum Glycophorin Binding Protein 130 Gene Promoter Activity[J]. , 2004, 22(2): 2-75.

[1]	LI Fei, LIU Yaobao, CAO Jun. Researches progress on molecular surveillance of artemisinin resistance in Plasmodium falciparum [J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2025, 43(2): 260-268.
[2]	LI Suhua, JI Penghui, WANG Dan, WANG Hao, ZHOU Ruimin. Analysis of polymorphism of Pfhrp2 gene in imported Plasmodium falciparum in Henan Province in 2020 [J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2024, 42(5): 664-667.
[3]	YAN He, HUANG Fang, FENG Jun, YIN Jianhai, XIA Zhigui, CAO Jianping. Polymorphism of sulfadoxine-pyrimethemine resistant gene of Plasmodium falciparum in China-Myanmar border area from 2010 to 2018 [J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2024, 42(2): 153-159.
[4]	ZHOU Ruimin, JI Penghui, LI Suhua, YANG Chengyun, LIU Ying, QIAN Dan, DENG Yan, LU Deling, ZHAO Yuling, ZHAO Dongyang, ZHANG Hongwei. Polymorphism analysis of drug resistance genes in imported Plasmodium falciparum isolates from Equatorial Guinea in Henan Province [J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2023, 41(5): 593-600.
[5]	XU Shaojie, CHEN Shenbo, CHEN Junhu. Research progress on transcription regulation of rif gene in Plasmodium falciparum [J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2023, 41(3): 374-379.
[6]	TIAN Bin, LIAO Yu, WEN Lan, XIAO Fang, ZHANG Bin, SHEN Xiao-jun. Analysis on the copy number variation of multidrug resistance-1 gene in 122 imported cases of falciparum malaria in Changsha [J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2022, 40(1): 127-131.
[7]	SHI Ming-li, XIAO Bo, JIANG Lu-bin. Research progress on the expression regulation of var genes in Plasmodium falciparum [J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2021, 39(6): 719-724.
[8]	LI Mei, TU Hong, XIA Zhi-gui, WANG Zhen-yu, ZHOU He-jun. Thermal stability of diagnostic targets Plasmodium falciparum histidine rich protein Ⅱ and Plasmodium lactate dehydrogenase in rapid detection [J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2021, 39(2): 245-248.
[9]	SHI Shan-mei, CHEN Jun-hu. Research progress on the structure and function of RIFIN protein of Plasmodium falciparum [J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2021, 39(2): 249-255.
[10]	ZHANG Cang-lin, NIE Ren-hua, XU Dan, LV Gao-wei, WANG Jian, YANG Ya-ming, DENG Yan, LIU Yan, ZHOU Hong-ning. Correlation of Pfcrt, Pfmdr and PfK13 gene polymorphisms and in vitro drug susceptibility of Plasmodium falciparum isolates from China-Myanmar border region [J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2020, 38(5): 580-588.
[11]	YE Sheng-yu, CHENG Yi-yi, LI Man, ZHOU Hong-ning. An overview on the resistance of Plasmodium falciparum to primary anti-malarial drugs in China [J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2020, 38(5): 631-636.
[12]	YE Sheng-yu, CHENG Yi-yi, LI Man, ZHOU Hong-ning. Advances in methods for detecting drug-resistance molecular markers of Plasmodium falciparum [J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2020, 38(4): 490-495.
[13]	Ren-qing LI, Xiao-mei WANG, Yu-lan SUN, Yan-ning LV, Xiang-feng DOU, Quan-yi WANG. Application of metagenomic Next-Generation Sequencing in the diagnosis of imported malaria [J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2019, 37(6): 727-729.
[14]	Yun-shan MOU, Lu-jie LI, Yin-juan WU, Xue-rong LI. Exploration of molecular mechanisms of artemisinin resistance in malaria parasites [J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2018, 36(6): 636-642.
[15]	Chun LIU, ALFRED Ndoumadiamba, Gou GNONDA Mounzie. Clinical application of colloidal-gold detection reagent for Plasmodium falciparum in Gabon of Africa [J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2018, 36(6): 679-680.

Effects of Tetracycline Operator Element Insertion on Plasmodium falciparum Glycophorin Binding Protein 130 Gene Promoter Activity

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments