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当前位置: 首页 > 文章 > 瑟氏泰勒虫p23与牛IL-18复合基因真核表达载体的构建与瞬时表达 中国兽医学报 2015,35 (6) 922-925
Position: Home > Articles > Constructing transient eukaryotic expression vector of the Theileria Sergenti p23 and cattle IL-18 genes Chinese Journal of Veterinary Science 2015,35 (6) 922-925

瑟氏泰勒虫p23与牛IL-18复合基因真核表达载体的构建与瞬时表达

作  者：
孙平杰;宋建臣;李海峰;许应天
单  位：
延边大学农学院
关键词：
瑟氏泰勒虫;p23基因;牛IL-18;串连基因;真核表达
摘  要：
为构建瑟氏泰勒虫p23-IL-18复合基因真核表达质粒,以pMD-18T-p23和pMD-18T-IL-18克隆质粒为模板,应用重组PCR技术得到瑟氏泰勒虫p23-IL-18复合基因,将其先克隆到pMD-18-T载体,再亚克隆到真核表达载体pVAXⅠ中,得到重组质粒pVAXI-P23-IL-18。对该重组质粒进行PCR、酶切鉴定及测序后,采用脂质体法将其转染到BHK-21细胞,用RT-PCR和IFA进行鉴定。结果表明,瑟氏泰勒虫p23-IL-18复合基因真核表达质粒pVAXIP23-IL-18构建成功,可在BHK-21细胞中表达。本试验为p23-IL-18复合基因的免疫学研究奠定了基础。
译  名：
Constructing transient eukaryotic expression vector of the Theileria Sergenti p23 and cattle IL-18 genes
作  者：
SUN Ping-jie;SONG Jian-chen;LI Hai-feng;XU Ying-tian;Agricultural College,Yanbian University;
关键词：
Theileria sergenti;;p23gene;;interleukin-18(IL-18)gene;;tandem gene;;eukaryotic expression
摘  要：
Two pair of primers were designed by Oligo.6according to the sequence reported p23 gene of Theileria sergenti and IL-18 gene of cattle in the GenBank.Theileria sergenti p23 gene was linked with cattle IL-18 gene by SOE-PCR technique and then inserted into pMD18-T simple vector.The positive plasmid was digested with restriction-endonuclease of BamHI and EcoRI,and then the fusion gene was inserted into the eukaryotic expression vector pVAX1,named as pVAXp23-IL-18,the plasmid was transfected into BHK-21 cells with liposome,the expression of the fusion protein were analyzed by RT-PCR and IFA.The result indicated that pVAX-p23-IL-18 had been transfected into HeLa cells and transiently expressed successfully,which established the foundation for further study of the IL-18 and p23recombination gene as elect component of vaccinum.

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