中国畜牧兽医2012，Vol.39Issue(3)：47-50,4.
鸡γ-干扰素基因的克隆及原核表达
Cloning and Expression of Chicken IFN-γ Gene
芦芝川 1史耀旭 2杨孝朴2
作者信息
- 1. 甘肃农业大学动物医学院,甘肃兰州 730070
- 2. 中农威特生物科技股份有限公司甘肃兰州 730046
- 折叠
摘要
Abstract
The total RNA was abstracted from the concanavalin A(Con A) -activated spleen lymphocytes, a pair of specific primers was designed by Oligo software based on the sequence of chicken interferon-gamma submitted by digby in GenBank (accession No. U27465). A 500 bp DNA fragment was amplified by reverse transcription-polymerase chain reaction (RT-PCR) and then ligated into T-easy vector for sequencing. The results showed that the fragment contained the complete open reading frame (ORF) of CHIFN-r gene. In comparison with GenBank data, the homology of the nucleotide sequence was 100%. The sequence was inserted into pGEX-4T-1 and the recombined expressing vector was constructed, then transformed into Escherichia coli BL21, which were further induced by IPTG and cultured,and a fusion protein was obtained with about 43 ku of molecular weight in SDS-PAGE. This results showed that the cloned CHIFN-y gene was expressed in prokaryotic cells.关键词
γ-干扰素/鸡/克隆/表达Key words
interferon-y/ chicken/ cloning/ expression分类
生物科学引用本文复制引用
芦芝川,史耀旭,杨孝朴..鸡γ-干扰素基因的克隆及原核表达[J].中国畜牧兽医,2012,39(3):47-50,4.
