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8814402(½ð±Ò+2): ¹ÄÀøÐÂÊÖ£¡»¶Ó­³£À´¡£ 2011-08-14 16:20:53
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We extracted RNA from 18 kinds of pig tissues separately£¬anti-transcripted them to cDNA£¬and used RT-PCR method to amplify the gene code aera sequences of tumor necrosis factor£¬then jointed the purified sequences with pMD18-T carrier to transform E.coli. Positive clones were screened and their sequences were determined.Using bio-information method to analyse their characters£¬system evolution tree was set up with other species.Meanwhile,we used semi-quantitative RT-PCR method to analyse the expression of TNF-¦Á gene in 18 kinds of pig tissues . The result showed the whole length of pig TNF-¦Á gene code aera was 699 bp£¨GenBank accession number£ºJF831365 £©£¬it coded 232 amino acids. The amino acid homology analysis showed it had over 80% homology with 12 species like cat£¬molecular system evolution research indicated it had a close relation with cattle£¬goat£¬sheep and dolphin.Multi-tissues semiquantitative RT-PCR study showed TNF-¦Á mRNA expressed in several tissues of pig£¬and its expression was higher in brain£¬ovary£¬spleen£¬lung£¬neuro and stomach.
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