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Cloning, Expression and Polymorphism Analysis of Porcine SRPK3 Gene |
E Guang-xin1, LIU Di2, ZHANG Dong-jie2, CUI Yu3 |
1. Chinese Academy of Agricultural Science, Institute of Animal Sciences, Beijing 100193, China;
2. Heilongjiang Academy of Agricultural Science, Harbin 150086, China;
3. Animal Science College, Hainan University, Haikou 570228, China |
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Abstract Protein SRPK3 acted as a crucial element of transcription pre-initiation complex, which play an important role in regulation procession of gene expression. In order to explore the genetic characteristic of SRPK3 in pigs. SRPK3 gene came from Yorkshire a pig was cloned by RT-PCR yet coding sequence was completed. The distribution determination of mRNA came from ten Yorkshire pigs and Duroc pigs in heart, muscle, liver, kidney, lung stomach, small and large intestine, spleen, brain was finished by Real-time PCR by age one day and 30 days. Expression test of gene SRPK3 was implemented in a skeletal damage model during the period of skeletal muscles development. Sequence analysis of an mRNA fragment with a length of 1 708bp in gene SRPK3 of a Yorkshire pig revealed a full coding region, 1 701bp, which coded 656 AAs including two S_TKc domains. The sequence of the porcine protein SRPK3 shared high similarity with its homolog from human and bovine, and they were closed in the Phylogenetic tree. PCR-SSCP test shows: On the Sixth exon (CDS: 629, CDS: 653) get A→G,T→C, which change the amino as Pro → His,Ile→Thr, on the ninth exon (CDS: 1059) get a G → A, but no change of amino. Both breeds specific expression and tissues specific expression were detected by RT-PCR, however high expression was mainly detected in skeletal muscle and heart. The quantity of mRNA of gene SPRK3 in the period of skeletal muscle destruction and repair, which showed gradually to be increased in different stage. Because of it’s expressed in skeletal muscle and heart mainly, and gradually be increased in different stage of injury and repair process.SRPK3 may be related to skeletal muscle cell development.
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Received: 28 July 2010
Published: 01 April 2011
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Fund: Supported by Eleventh Five-Year Plan Key Projects National Science and Technology(2008BADB2B02) |
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