[The expression and analysis of its activity of anti-bacterial peptide gloverin in COS-7 cells].
OBJECTIVE
To investigate the expression and analysis of its activity of anti-bacterial peptide gloverin in COS-7 cells.


METHODS
The appearance frequency of all genetic codes in the cDNA sequence from the same species of protein Attacin A was analyzed, and its cDNA sequence was synthesized by PCR overlapping extension method in conjunction with the designation of the known protein sequence of gloverin. The genes were inserted into pCDSI, an eukaryotic vector, after being identified correctly. As a result, the vector pBZHG was constructed. Thereafter, the liposome FuGENE( trade mark ) 6 was employed as the vector, and the COS-7 cells were transfected with liposome pBZHG and blank vector pCDSI. The normal cells were taken as the control. The supernatant was collected for the detection of its bactericidal activity after 72 PBHs.


RESULTS
The gloverin cDNA sequence designed artificially was expressed in COS-7 cells. The supernatant of the cells transfected by pBZHG exhibited bactericidal activity to E. coli J5 when compared with that from normal cells and in cells transfected with blank vectors.


CONCLUSIONS
The designed cDNA sequence of gloverin was proved to be genuine, and it provided the basis for future study of its antibiotic and anti-endotoxin activities.
Zhou
Hong
H
Department of Pharmacology, College of Medicine, The Third Military Medical University, Chongqing, 400038. P.R. China.
Zheng
Jiang
J
Lu
Yong-Ling
YL
Qin
Xiao-Jian
XJ
Xiao
Guang-Xia
GX
D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D019556:COS Cells; D002522:Chlorocebus aethiops; D036341:Intercellular Signaling Peptides and Proteins; D011506:Proteins / Q000235:genetics* / Q000494:pharmacology; D017422:Sequence Analysis, DNA; D014162:Transfection
D000900:Anti-Bacterial Agents; D036341:Intercellular Signaling Peptides and Proteins; D011506:Proteins; C107617:gloverin