›› 2011, Vol. 38 ›› Issue (1): 95-99.

• 生物技术 • Previous Articles     Next Articles

Cloning and Sequence Analysis of the α-galactosidase Gene (melA)from L. plantarum

REN Da-yong1,2,3,LI Chang2,QIN Yan-qing2,3,DU Shou-wen1,2,HU Bo1,2,JIN Ning-yi2   

  1. (1.College of Animal and Veterinary Medicine,Jilin University, Changchun 130062, China;2.Military Veterinary Institute, Academy of Military Medical Sciences, Changchun 130062, China;3.College of Food Science and Engineering, Jilin Agricultural University, Changchun 130118, China)
  • Received:1900-01-01 Revised:1900-01-01 Online:2011-01-01 Published:2011-01-01
  • Contact: JIN Ning-yi

Abstract: Using chromosome DNA of L. plantarum as template,melA gene was amplified by PCR and purified by purification kit in this study. The PCR product of melA gene was cloned into pMD18-T vector and transformed into DH5α competent cells. The recombinant plasmid containing melA gene was selected and the inserted melA gene was identified by cleaved with restriction endonuclease Sac Ⅰ and Sph Ⅰ, and following by PCR amplification and sequencing. The cloned complete melA gene sequence, at a length of 2240 bp, encoded 738 amino acids with a deduced molecular weight of 84 ku. The GC and AT contents were 47.45% and 52.55% respectively. Three nucleotides nonsense mutation were observed, resulting no change of deduced amino acids. The homologies of nucleotides and amino acids were more than 99% respectively (except for AY873840, 96.6% and 92.2%) compared with those in GenBank. The results showed that the whole melA gene was cloned successfully. This research proved a foundation for the construction of recombinant express vector of melA gene.

Key words: L. plantarum; α-galactosidase; cloning; sequence analysis

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