Construction, expression and biological assessment of BPI23-Fcgamma1 recombinant protein prokaryotic expression vector / 中国医学科学杂志(英文版)
Chinese Medical Sciences Journal
;
(4): 140-147, 2002.
Article
in English
| WPRIM
| ID: wpr-244861
ABSTRACT
<p><b>OBJECTIVE</b>To construct pBV-BPI600-Fcgamma1(700) recombinant expression vector, to transform it into Escherichia coli DH5alpha, and to induce the expression of BPI23-Fcgamma1 anti-bacterial recombinant protein.</p><p><b>METHODS</b>Genes coding for BPI23 and Fcgamma1 were amplified by RT-PCR from mRNA extracted from HL-60 cell and normal human leukocytes; recombinant cloning vector and recombinant expression vector were then constructed. pBV-BPI600-Fcgamma1(700) recombinant expression vector was transformed into the competent Escherichia coli DH5alpha and BPI23-Fcgamma1 recombinant protein was expressed by a temperature-induced method.</p><p><b>RESULTS</b>(1) Expected amplified products BPI600hp and Fcgamma1(700bp) were obtained by RT-PCR method. (2) pUC18-BPI180, pUC18-BPI420 and pUC18-Fcgamma1(700) recombinant cloning vector were successfully constructed, and sequences were identical with the reported ones. 3) pBV-BPI600-Fcgamma1(700) recombinant expression vector was successfully constructed, and the enzyme digestion analysis showed an expected result. (4) The expression level of BPI23-Fcgamma1 recombinant protein accounted for 20% of total bacterial proteins. (5) The renatured BPI23-Fcgamma1 recombinant protein showed bacteriocidal activity and biological function of complement fixation, and opsonization.</p><p><b>CONCLUSION</b>pBV-BPI600-Fcgamma1(700) recombinant expression vector was successfully constructed, and BPI23-Fcgamma1 recombinant protein with double biological activity of BPI and IgGFc was expresed in Escherichia coli.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Pharmacology
/
Blood Bactericidal Activity
/
Recombinant Fusion Proteins
/
Blood Proteins
/
Carrier Proteins
/
Cell Adhesion Molecules
/
HL-60 Cells
/
Antimicrobial Cationic Peptides
/
Escherichia coli
/
Genetic Vectors
Limits:
Humans
Language:
English
Journal:
Chinese Medical Sciences Journal
Year:
2002
Type:
Article
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