Expression of Human Interferon α 2b Gene in Ginseng Cells  

作　　者：REN Qi WANG Chun-yi SONG Zhi-ming LIU Dan YU Hai-peng SHENG Jun 

机构地区：[1]College of Life Science, Jilin University, Changchun 130012, P. R. China [2]Changchun Institute of Biological Products, Changchun 130062, P R. China [3]Liaoning Yisheng Biopharmaceutical Co., Ltd., Shengyang 110131, P. R. China [4]First Affiliated Hospital, Jilin University, Changchun 130031, P R. China [5]Yunnan Agricultural University, Kunming 650201, P.. R. China

出　　处：《Chemical Research in Chinese Universities》2010年第3期420-426,共7页高等学校化学研究（英文版）

基　　金：Supported by the Science and Technology Development Planning Foundation of Jilin Province of China(No.20030405)

摘　　要：The human interferon α 2b（hIFN-α2b） gene was cloned into binary vector pBI121 to obtain plant expression vector pBIFN. The recombinant plasmid pBIFN was transferred into Agrobacterium tumefaciens strain LBA4404. Then the hIFN-α2b gene was introduced into ginseng callus cells via Agrobacterium-mediated transformation and the ginseng cell line carrying hIFN-α2b gene was selected on G418-containing medium. The presence of target gene in transformed cells was confirmed by PCR and RT-PCR. The results indicate that hIFN-α2b gene has been integrated into the ginseng cells＇ genome, with transcription products, hIFN-α2b expressed by the transgenic ginseng cells was detected by Western blot. It was shown that a specific protein band at 19000 could be observed. Cytopathic effect（CPE） inhibition assay using the W1SH-VSV system shows that the mean antiviral activity of expressed hlFN-a2α was 6.0× 10^4 IU/mL.The human interferon α 2b（hIFN-α2b） gene was cloned into binary vector pBI121 to obtain plant expression vector pBIFN. The recombinant plasmid pBIFN was transferred into Agrobacterium tumefaciens strain LBA4404. Then the hIFN-α2b gene was introduced into ginseng callus cells via Agrobacterium-mediated transformation and the ginseng cell line carrying hIFN-α2b gene was selected on G418-containing medium. The presence of target gene in transformed cells was confirmed by PCR and RT-PCR. The results indicate that hIFN-α2b gene has been integrated into the ginseng cells＇ genome, with transcription products, hIFN-α2b expressed by the transgenic ginseng cells was detected by Western blot. It was shown that a specific protein band at 19000 could be observed. Cytopathic effect（CPE） inhibition assay using the W1SH-VSV system shows that the mean antiviral activity of expressed hlFN-a2α was 6.0× 10^4 IU/mL.

关 键 词：Human interferon × 2b Ginseng cell Plant transformation Agrobacterium tumefaciens 

分 类 号：S567.51[农业科学—中草药栽培] S436.421[农业科学—作物学]