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中国生物工程杂志

China Biotechnology
China Biotechnology
China Biotechnology  2011, Vol. 31 Issue (7): 27-31    DOI:
    
Construction and Verification of Skeletal Muscle Specific Expression Vector of Caenorhabditis Elegans ω-3 Fatty Acid Desaturase Gene
YU Yong-sheng1,2, ZHANG Li-chun2, LUO Xiao-tong2, LIU Zheng2, ZHANG Shu-min2
1. Northeast Agricultural Research Center of China, Changchun 130124, China;
2. Branch of Animal Husbandry, Jilin Academy of Agricultural Sciences, Gongzhuling 136100, China
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Abstract  

Two specific oligonucleotide primers were designed and synthesized according to the sequence of Sus scrofa skeletal α-actin gene, which were used to amplify the α-actin 5'regulating sequence. pcDNA3.1-AF, the muscle-specific expression vector, was constructed by connecting with α-actin 5'regulating sequence, Caenorhabditis elegans ω-3 fatty acid desaturase cDNA and CMV promoter-less pcDNA3.1,and its expression specificity were proved by directly injecting the vector into the quadriceps muscle of mice(4 weeks old). The PCR amplification result showed that a size of 1 815 base pairs fragment was amplifies and the sequencing resulted showed that the fragment had a high identity with original sequence and contained muscle specific elements. The transient expression results indicated that the muscle-specific expression vector could efficiently direct the transcription of ω-3 fatty acid desaturase in the muscle. The study provides the basis for establishing transgenic animal which could specific express ω-3 fatty acid desaturase in the muscle.

Key words： Sus scrofa skeletal α-actin      Caenorhabditis elegans ω-3 fatty acid desaturase      Muscle-specific expression vector      Verification in vitro     
Received: 03 February 2011      Published: 25 July 2011
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YU Yong-sheng
ZHANG Li-chun
LUO Xiao-tong
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ZHANG Shu-min
Cite this article:

YU Yong-sheng, ZHANG Li-chun, LUO Xiao-tong, LIU Zheng, ZHANG Shu-min. Construction and Verification of Skeletal Muscle Specific Expression Vector of Caenorhabditis Elegans ω-3 Fatty Acid Desaturase Gene. China Biotechnology, 2011, 31(7): 27-31.

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https://manu60.magtech.com.cn/biotech/     OR     https://manu60.magtech.com.cn/biotech/Y2011/V31/I7/27


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