[Cloning and eukaryotic expression of the gene encoding myosin from Brugia malayi].
Total RNA was extracted from periodic microfilariae of Brugia malayi and its myosin partial gene (Bm-M55) was amplified by RT-PCR. The PCR product was cloned and then subcloned into pcDNA3.1 (+)vector. The recombinant eukaryotic plasmids were screened and identified by digestion with restriction enzyme and PCR amplification, and was transfected into COS-7 cells subsequently. The expressed protein was identified by SDS-PAGE. Bm-M55 mRNA was highly expressed in transfected COS-7 cells. The deduced amino acid sequence showed to be identical with that of Bm-M55, and the recombinant protein was about Mr 55000.
Xie
Dong-fang
DF
Department of Parasitology, School of Basic Medical Sciences, Nantong University, Nantong 226001, China.
Fang
Zheng
Z
Huang
Wei-qun
WQ
Shen
Qin
Q
Tong
Hai-yan
HY
Xu
Bang-sheng
BS
D000818:Animals; D017178:Brugia malayi / Q000235:genetics* / Q000378:metabolism; D019556:COS Cells; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D015870:Gene Expression; D017238:Genes, Helminth; D005822:Genetic Vectors; D009218:Myosins / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
D009218:Myosins