›› 2012, Vol. 39 ›› Issue (3): 47-50.

• 生物技术 • Previous Articles     Next Articles

Cloning and Expression of Chicken IFN-γ Gene

LU Zhi-chuan1,2, SHI Yao-xu2, YANG Xiao-pu1   

  1. 1. College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070,China;2. China Agriculture Vet.Bio.Science and Technology Co., Ltd , Lanzhou 730046, China
  • Received:2011-11-01 Revised:1900-01-01 Online:2012-03-20 Published:2012-03-20

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-γ 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-γ gene was expressed in prokaryotic cells.

Key words: interferon-γ; chicken; cloning; expression

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