[Cloning and identification of human vigilin full-length coding region].
OBJECTIVE
The full-length of human vigilin encodes 1289 amino acids, containing 14 KH domains. We adopted a five step PCR technique to clone the full-length vigilin coding regions to investigate the functions of full-length vigilin, N-terminal of vigilin, C-terminal of vigilin and different KH.


METHODS
Vigilin was dissected into five fragments according to the restrictive endounclease enzymes site of vigilin. The total RNA was extracted from the livers of healthy adults died from accident. The cDNA was amplified by RT-PCR. Then the amplified PCR product was digested and inserted into pUC118 vector and subcloned into pUC118 vector.


RESULTS
Eight gene fragments and full-length of human vigilin were obtained. The gene sequence of vigilin was identical with its cDNA sequence reported (NM:005336) in the GenBank.


CONCLUSIONS
The full-length vigilin coding region has been cloned successfully.
Yang
Wen-li
WL
Department of Biochemistry and Molecular Biology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China.
Wu
Jing
J
Xie
Xiao-yan
XY
Wei
Ling
L
Qin
Yang
Y
D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D006801:Humans; D016366:Open Reading Frames / Q000235:genetics*; D016601:RNA-Binding Proteins / Q000032:analysis / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods
D018076:DNA, Complementary; D016601:RNA-Binding Proteins; C075197:high density lipoprotein binding protein