OBSTRUCTION AND SEQUENCE DETERMINATION OF E.COLI-MYCOBACTERIA RECOMBINANT SHUTTLE PLASMID OF CSP GENE FRAGMENT OF PLASMODIUM FALCIPARUM
10.3969/j.issn.1005-0507.2001.01.002
- VernacularTitle:恶性疟原虫FCC-1/HN株环子孢子蛋白基因分枝杆菌——大肠杆菌穿梭表达重组质粒的构建及序列测定
- Author:
Chunfu ZHENG
1
;
Shaoting WU
;
Yatang CHEN
;
Shitong GAO
;
Min LIN
Author Information
1. 重庆医科大学附属第一医院
- From:
Acta Parasitologica et Medica Entomologica Sinica
2001;8(1):7-12
- CountryChina
- Language:Chinese
-
Abstract:
The circumsporozoite protein(CSP) gene fragment of Plasmodium falciparum was cloned and sequenced.A pair of primer was designed according to the CSP encoding sequence of 837 isolate(Thailand isolate),and the CSP gene fragment was amplified by polymerase chain reaction(PCR) from Plasmodium falciparum FCC-1/HN isolate,it spanned the conserved region I.the central immunodominant repeat region\,the variable region behind the repeats and the conserved II region.After purification,the CSP gene fragment was digested with restriction enzyme BamH I and Kpn I,and ligated with the pBCG5.6 digested with the same enzyme.The recombinant pBCG5.6/CSP was transformed into E.coli DH5α.Positive clones were screened and identified by PCR technique and digestion with restriction enzyme.Nucleotide sequence was determined by dideotide chain-termination method.The results indicated that the CSP gene was successfully amplified and cloned,its nucleotide sequence was about 1 171 bp and was in accordance with the expected one.Sequence determination results showed that the cloned gene fragment was the same with the CSP encoding gene.
