当前位置: 首页 > 文章 > 鸡IL-2全基因和去信号肽基因的酵母表达研究 郑州牧业工程高等专科学校学报 2009,29 (3) 1-6
Position: Home > Articles > The P.pastoris Expression of ChIL-2 whole gene and partial gene without singlnal sequence Journal of Zhengzhou College of Animal Husbandry Engineering 2009,29 (3) 1-6

鸡IL-2全基因和去信号肽基因的酵母表达研究

作  者:
龚婷;李银聚
单  位:
河南科技大学动物科技学院;郑州牧业工程高等专科学校生物工程系
关键词:
鸡IL-2;信号肽;酵母表达;mRNA二级结构
摘  要:
根据已发表的鸡白介素-2(ChIL-2)cDNA编码基因序列设计三条引物,利用RT-PCR技术从经诱导的卢氏鸡胚脾淋巴细胞中扩增出429bp的ChIL-2全长基因cDNA和363bp去除信号肽的ChIL-2基因cDNA。分别克隆到酵母表达载体pPICZαA中,并分别转化巴斯德毕赤酵母X-33中,经甲醇诱导,SDS-PAGE检测,在28kD和24kD处有两条清晰的蛋白带,相同条件下pPICZαA-△SIL-2表达量较高。RNAstructure软件分析表明,pPICZαA-IL-2的序列形成了较为稳定的二级结构,不利于翻译,证明信号肽序列的存在对ChIL-2基因在酵母细胞中的表达有一定的影响。
译  名:
The P.pastoris Expression of ChIL-2 whole gene and partial gene without singlnal sequence
作  者:
Gong Ting1,Li Yin-ju2(1.Biology Engineering Ddepartment,Zhengzhou College of Animal Husbandry Engineering,Zhengzhou,Henan 450011; 2.Animal and Science College,Henan University of Science and Technolygy,LuoYang,471003)
关键词:
Chicken interleukin-2;Signal peptide;Yeast express,mRNA Secondary structure
摘  要:
Three primers were designed according to the ChIL-2 gene sequences in the GenBank and the whole 429bp gene and 363pb gene without signal sequence of ChIL-2 were amplified from the induced spleen lymph cells of Lushi chicken embryo by using RT-PCR,which were cloned into the P.pastoris express vector pPICZαA respectively to construct the P.pastoris recombinate plasmid pPICZαA-IL-2,pPICZαA-△SIL-2.and then transformed to P.pastoris X-33.IL-2 was expressed under the induction of methanol.There were two clear protein band in the 28 kD and 24 kD,which were identified by SDS-PAGE.Under the same conditions pPICZαA-△SIL-2 expression was higher.RNAstructure software analysis showed that sequences of pPICZαA-IL-2 formed a complex and relatively stable secondary structure.It is not conducive to translation which testifies that the existence the signal peptide sequence has influence to ChIL-2 gene in P.pastoris express.

相似文章

计量
文章访问数: 11
HTML全文浏览量: 0
PDF下载量: 0

所属期刊

推荐期刊