Cloning of Porcine IL-2 Gene and Its Construction of Recombinant Fowlpox Viruses Vector

doi:10.7668/hbnxb.2011.05.015

Abstract

Abstract: Porcine IL-2 gene was amplified by RT-PCR from the total RNA extracted from the splenocyte of Yu′nan pig.The PCR product was cloned and sequenced.The porcine IL-2(pIL-2)gene was digested by BamHⅠand cloned into pSY538 vector which was digested by BamHⅠand dephosphorized by calf alkaline phosphatase.By means of the PCR,restriction enzyme digestion and sequencing,the recombinant plasmid pSY538/pIL-2 was selected.After pSY538/pIL-2 was digested by NotⅠ,the pIL-2 fragment containing LP2 and EP2 was achieved and subcloned into pSY681 vecter containing LacZ gene,then the recombinant plasmid pSY681/pIL-2 was developed.The results indicated that the cloned pIL-2 gene had 482 bp,including one ORF(465 bp).The IL-2 gene sequence of Yu′nan pig had 99.9% nucleotide homology with the 5 pig IL-2 genes published in GenBank.Enzyme digestion and DNA sequencing results confirmed that the sequence of the recombinant plasmid pSY681/pIL-2 contained the target fragment,and the ligation part was correct.The results showed that the recombinant plasmid pSY681/pIL-2 was constructed correctly,paving the way for further study of biological function and further application of porcine IL-2.

Key words: Porcine IL-2, Clone, Recombinant fowlpox virus, Vector, Construction

CLC Number:

S828.2

WANG Shu-juan, CHEN Hong-ying, WEI Zhan-yong, LIU Jin-peng, GENG Jing-wei, CUI Bao-an. Cloning of Porcine IL-2 Gene and Its Construction of Recombinant Fowlpox Viruses Vector[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2011, 26(5): 71-75. doi: 10.7668/hbnxb.2011.05.015.

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