摘要
将荣昌猪IFN-ω基因亚克隆到酵母表达载体pPICZαB后,氯化锂法转化毕赤酵母菌株X-33,通过酶切、PCR鉴定表明成功构建了表达荣昌猪IFN-ω基因的酵母基因工程菌。经甲醇诱导后,通过SDS-PAGE和Western blotting检测,表明荣昌猪IFN-ω基因在毕赤酵母中实现了高效表达。表达产物在MDBK细胞上抗VSV的比活性为1.21×106U/mg,表明具有抗病毒活性。
The IFN-ω gene of Rongchang swine was subcloned to yeast expression vector pPICZαB.The recombinant plasmid was transformed into Pichia pastoris X-33 by LiCl after linearised with SacⅠ,after which the recombinant was identified by PCR and digestion with restriction enzymes.The vector with IFN-ω of Rongchang swine was induced with methanol for 4 days.With detection of SDS-PAGE and Western blotting,it was found that the target protein was highly expressed in Pichia pastoris.The antiviral activity of the purified product was tested by CPE50 method,which was 1.21×106 U/mg against VSV in MDBK cells.It was shown that the recombinant protein of IFN-ω had antiviral activity.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2010年第12期1602-1605,共4页
Chinese Journal of Veterinary Science
基金
西南大学青年基金资助项目(QNRC200905)
