CONSTRUCTION OF cDNA LIBRARY OF TOXOPLASMA GONDII

Abstract

Abstract: ATM: To construct a cDNA library of Toxoplasma gondii. METHOD: We extracted the RNA of Toxoplasma gondii (RH strain)by using a single step acid guanidinium thiocyanate method and isolated the poly(A) +RNA by using messenger RNA affinity membrane containing poly U to synthesize ds cDNAs.After ligating the cDNA into the adaptor of EcoRI/NotI and then into λgt11 DNA arms of the expressive vector,the ligated products were packaged in vitro and used to infect the E.coli Y 1090 .The 32 P dCTP genomic DNA of T.gondii RH strain hybridized the λgt11 recombinant plaques in situ .RESULTS:The ds cDNAs were found to be 0.5-2 kb in size.The cDNA library of T.gondii constructed showed 6.97×10 5 recombinant plaqes.The recombination rate and the cloning efficiency were 98.73% and 6.97×10 6 clones/μg cDNA,respectively.For in situ hybridization of the λgt11 recombinant plaques,the rate of hybridization was 95.2%.CONCLUSION:The method of extracting the RNA of T.gondii is simple and efficient.The cDNA library constructed has large content and good quality.

CLC Number:

R382.5

[1]	JING Tianyu, MOU Rutao, ZHANG Fan, LIU Xianbing, HU Xuemei, ZHANG Haixia, LI Zhidan. Effect of Toxoplasma gondii infection during pregnancy on CD73 expression in trophoblasts and decidual immune cells and the association of CD73 expression with adverse pregnancy [J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2025, 43(2): 210-216.
[2]	ZHANG Xiaocheng, HU Yuan, PENG Hui, SHEN Yujuan, LIU Hua, CAO Jianping. CRISPR/Cas9-based generation of the bfd2 deficient strain of Toxoplasma gondii and analysis its phenotype [J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2025, 43(2): 217-222.
[3]	YIN He, MA Lei, DANG Tiantian, LI Jiaming, Zhao Zhijun. Effect and mechanism of Toxoplasma gondii ROP16 Ⅰ/Ⅲ regulating TAF15 on THP-1 cells [J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2025, 43(1): 103-111.
[4]	LI Shiyu, LI Jing, LU Shaohong, ZHENG Bin. Research advances on host cell-autonomous immunity against Toxoplasma gondii [J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2024, 42(5): 653-658.
[5]	XIE Xiaoman, SUN Hang, DAI Lisha, ZHU Wenju, WANG Lilei, XIE Huanhuan, DONG Hongjie, ZHANG Junmei, WANG Qi, ZHOU Beibei, ZHAO Guihua, XU Chao, YIN Kun. Effect of Toxoplasma gondii infection on m⁶A methylation modification of transcripts in mice brain tissue [J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2024, 42(1): 27-35.
[6]	XUE Yushan, LIN Ping, CHENG Xunjia, FENG Meng. Damage caused by chronic infection of Toxoplasma gondii on the host central nervous system and its mechanism [J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2023, 41(5): 527-531.
[7]	JIANG Wenjing, MENG Yali, ZHAO Lina, WANG Chunmiao, ZHANG Xiaolei. Immunoprotection of nuclei acid vaccine dual-targeting rhoptry protein 18 and surface antigen 30 of Toxoplasma gondii in mice [J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2023, 41(5): 532-538.
[8]	ZHAO Ziqi, LV Fangli. Study on the inhibitory effect of artemether liposome on Toxoplasma gondii proliferation in vitro [J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2023, 41(4): 446-451.
[9]	ZHANG Chi, CHEN Jiating, XIN Zixuan, YANG Lili, YANG Zihan, PENG Hongjuan. Transcriptome analysis of mice brain chronically infected with Toxoplasma gondii and validation of the kynurenine pathway associated with depression [J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2023, 41(3): 270-278.
[10]	LI Jia-ming, WANG Yi-xuan, YANG Ning-ai, MA Hui-hui, LAN Min, LIU Chun-lan, ZHAO Zhi-jun. Effects of ROP16 protein of Toxoplasma gondii on polarization and apoptosis of MH-S cells and their related mechanisms [J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2022, 40(5): 579-586.
[11]	ZOU Wei-hao, WU Wei-ling, LIAO Yuan-peng, CHEN Min, PENG Hong-juan. Preparation and application of monoclonal antibody against Toxoplasma gondii bradyzoite antigen 1 [J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2022, 40(5): 587-593.
[12]	WANG Jie, WEN Hong-yang, CHEN Ying, AN Ran, LUO Qing-li, SHEN Ji-long, DU Jian. Construction and identification of macrophage migration inhibitory factor gene knockout strain of Toxoplasma gondii [J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2022, 40(3): 349-354.
[13]	ZHANG Xiao-han, FENG Ying, CHEN Ran, SANG Xiao-yu, YANG Na. Advances in research of structure, function and regulatory mechanism of Toxoplasma gondii conoid [J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2021, 39(6): 832-835.
[14]	SU Ya-jing, QIAO Xia, WANG Peng-tao, KANG Yu-ting, YANG Ning-ai, JIA Wei, ZHAO Zhi-jun. Gene expression profiling of human lung adenocarcinoma of A549 cells induced by Toxoplasma virulence-related effector ROP16Ⅲ [J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2021, 39(4): 473-480.
[15]	WANG Long-jiang, LI Jin, YIN Kun, XU Chao, LIU Gong-zhen, HUANG Bing-cheng, WEI Qing-kuan, SUN Hui. Comparative analysis of transcriptomes in Toxoplasma gondii before and after invasion in human foreskin fibroblasts [J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2021, 39(4): 480-486.

CONSTRUCTION OF cDNA LIBRARY OF TOXOPLASMA GONDII

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments