[Expression and character of measles virus hemagglutinin gene].
OBJECTIVE
To express the hemagglutinin gene of measles virus (MV) in Escherichia col iexpression system, and apply the recombinant protein for detection of MV.


METHODS
Measles virus genomic RNA was extracted from the strain MVi/Zhejiang. CHN/7.05/4, the interest H gene was amplified by RT-PCR and subcloned into the pET28a (+) vector, the recombinant plasmid was transformed into E. coli BL21(DE3) cells and induced with IPTG. After the expression of the recombinant protein, it was analyzed with SDS-PAGE, while Western blotting was performed to determine the specific antigenicity of the target protein. Then, the recombinant protein purified with the Ni-NTA column was applied for the ELISA detection.


RESULTS
The interest gene amplified by RT-PCR was 1700bp, and there was no mutation by sequencing. The interest protein was expressed in inclusion in Escherichia coli expression system, and showed the high antigenicity in Western blotting.


CONCLUSIONS
The interest protein was successfully expressed in prokaryotic system, and can be used as the antigen in measles virus detection after it was purified.
Liang
Liang
L
The College of Life Science, Zhejiang Sic-Tech University, Hangzhou 310018, Zhejiang, China.
Chen
Yin
Y
Lu
Yi-Yu
YY
D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006388:Hemagglutinins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D008459:Measles virus / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D008970:Molecular Weight; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology
D006388:Hemagglutinins; D011994:Recombinant Proteins; D014764:Viral Proteins