Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (50): 9437-9440.doi: 10.3969/j.issn.1673-8225.2011.50.031

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Plasmid construction and variation analysis of exon-4 of carnitine palmitoyltransferase-Ⅱ gene

Yao Deng-bing1, Yao Min2, Wang Li2, Dong Zhi-zhen3, Yao Deng-fu4   

  1. 1Key Laboratory of Neuroregeneration, Nantong University, Nantong  226001, Jiangsu Province, China
    2Center of Medical Examination, Affiliated Hospital of Nantong University, Nantong  226001, Jiangsu Province, China
    3Department of Diagnostics, School of Medicine, Nantong University, Nantong  226001, Jiangsu Province, China
    4Research Center of Clinical Medicine, Affiliated Hospital of Nantong University, Nantong  226001, Jiangsu Province, China
  • Received:2011-06-01 Revised:2011-07-16 Online:2011-12-10 Published:2011-12-10
  • Contact: Yao Deng-fu, Doctor, Professor, Research Center of Clinical Medicine, Affiliated Hospital of Nantong University, Nantong 226001, Jiangsu Province, China yaodf@ahnmc.com
  • About author:Yao Deng-bing☆, Doctor, Associate professor, Key Laboratory of Neuroregeneration, Nantong University, Nantong 226001, Jiangsu Province, China yaodb@ntu.edu.cn Dong Zhi-zhen, Associate researcher, Department of Diagnostics, School of Medicine, Nantong University, Nantong 226001, Jiangsu Province, China dongzz@ahnmc. com Yao Deng-bing and Dong Zhi-zhen contributed equally to this paper.
  • Supported by:

    Natural Science Foundation of Nantong University, No. 09ZY014*,  09B16*, 09ZY007*; Natural Science Foundation for Colleges and Universities in Jiangsu Province, No. 09KJB310012*; Natural Science Foundation of Jiangsu Province, No. BK2010282*

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Abstract:

BACKGROUND: Carnitine palmitoyltransferase-Ⅱ (CPT-Ⅱ) is located in the inner mitochondrial membrane and is one of the pivotal enzymes during the fatty acid oxidation. Currently, there are no studies about influenza virus infection and variation of CPT-Ⅱ gene.
OBJECTIVE: To construct the plasmid expressing exon-4 of CPT-Ⅱ gene (CPT-Ⅱ-E4) and analyze its mutation.
METHODS: Human CPT-Ⅱ DNA from the blood of two patients infected with influenza virus was involved. After the PCR amplification, the product of CPT-Ⅱ-E4 was cloned into pGEMD-T vector. The T-CPT-ⅡE4 was transferred to E. coli DH5α cells after prepared with T4 DNA higase. Then propagation of bacteria, preparation of plasmid, EcoRI digestion, sequencing, and variation analysis were performed.
RESULTS AND CONCLUSION: The recombinant pGEM-T-CPT-ⅡE4 was proved to contain whole sequence of CPT-Ⅱ-E4 with 1305 nucleotides which could code 435 amino acids. According to the original sequence from Genebank, two variable sites were found at 1618 (GTC→ATC) and 1858 (TTT→TCT) which code amino acids of V368I and F448L respectively.

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