Yan Yu-Lan, Liu Yang, Cao Wen-Yan, Bu Xue-Feng, Bu Zhi-Gao, Zheng Jin-Xu
Basic Medicine and Medical Telenology College, Jiangsu University, Zhenjiang 212013, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2008 Oct;24(10):950-3.
To construct the eukaryotic expressing vector PCAGG -HuIFN-lambda1 and PCAGG-HuIFN-lambda2 and to study the biological activity of HuIFN-lambda1and HuIFN-lambda2.
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.
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.
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.
