[Human lamada-interferon expressed in BHK-21 cell line and its bioactivity].
OBJECTIVE
To construct the eukaryotic expressing vector PCAGG -HuIFN-lambda1 and PCAGG-HuIFN-lambda2 and to study the biological activity of HuIFN-lambda1and HuIFN-lambda2.


METHODS
The cDNA fragment encodding HuIFN-lambda1 and HuIFN-lambda2 was amplified from the total RNA extracted from virus-induced HeLa cells by RT-PCR. Then it was cloned into the eukaryotic expressing vector PCAGG-EGFP. The recombinant was transfected into BHK-21 cells. VSV*GFP-A549 system was used to measure the anti-virus activity.The constructed cell line MDBK-Mxp-Luc was used to study the characteristics of MxA protein induced by the products of PCAGG-HuIFN-lambda1 and PCAGG-HuIFN-lambda2.


RESULTS
The recombinant vector HuIFN-lambda1-PMD18-T Vector was enzymed by Sac I and Xho I while HuIFN-lambda2-PMD18-T Vector was enzymed by Sac I and Sal I. The fragments were both 610 bp and they were consistent with nucleotide sequences reported in GenBank. The anti-virus activity of protein expressed by PCAGG-HuIFN-lambda1 and PCAGG-HuIFN-lambda2 was 10(4) IU/mL and 10(2) IU/mL, respectively. The protein expressed by PCAGG-HuIFN-lambda1 and PCAGG-HuIFN-lambda2 induced the expression of the anti-virus protein MxA. The expression of protein MxA induced by PCAGG-HuIFN-lambda1 increased with the passage of time, reaching the peak during 9 to 12 hours and disappearing in 24 hours.


CONCLUSIONS
The eukaryotic expressing vector of PCAGG-HuIFN-lambda1 and PCAGG-HuIFN-lambda2 has been successfully constructed and transiently expressed in BHK-21 cells. The anti-virus activity of the products is closely correlated with inducing the expression of anti-virus protein MxA.
Yan
Yu-Lan
YL
Basic Medicine and Medical Telenology College, Jiangsu University, Zhenjiang 212013, China. ylyan2005@yahoo.com.cn
Liu
Yang
Y
Cao
Wen-Yan
WY
Bu
Xue-Feng
XF
Bu
Zhi-Gao
ZG
Zheng
Jin-Xu
JX
D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D018076:DNA, Complementary / Q000235:genetics; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics / Q000502:physiology; D006367:HeLa Cells; D006801:Humans; D007372:Interferons / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D008856:Microscopy, Fluorescence; D064547:Myxovirus Resistance Proteins; D020133:Reverse Transcriptase Polymerase Chain Reaction
D018076:DNA, Complementary; C579071:MX1 protein, human; D064547:Myxovirus Resistance Proteins; D007372:Interferons; C034510:CTCGAG-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific; D019204:GTP-Binding Proteins