Cloning and expression of porcine SRPK1 gene
Protein SRPK1 acts as a crucial element in the pre-initiation complex of transcription, which play an important role in the regulation procession of gene expression. This study was carried out in order to explore the genetic characteristic of SRPK1 in pigs. SRPK1 gene came from Yorkshire, a pig that was cloned by real time polymerase chain reaction (RT-PCR), yet coding sequence and partial 5’UTR sequence was completed. The distribution determination of mRNA taken from the heart, muscle, liver, kidney, lung stomach, small and large intestine, spleen and brain of ten Yorkshire and Duroc pigs was finished by real-time PCR by one day and 30 days old pigs. Expression test of gene SRPK1 was implemented in a skeletal damage model during the period of skeletal muscles development. Sequence analysis of a DNA fragment with a length of 2499 bp in gene SRPK1 of a Yorkshire pig revealed a full coding region that coded 656 AAs, yet including partial 5’UTR sequence. A total of 14 transcription binding sites were detected by bioinformatics analysis. Both the breeds’ specific expression and the tissues’ specific expression were detected by RT-PCR; however, high expression was mainly detected in the stomach, and the small and large intestines. The quantity of the mRNA of gene SPRK1 was enhanced and it decreased repeatedly in the period of skeletal muscle destruction and repair. All the results in the foregoing indicated a few transcription binding sites, which showed that the relevant muscle growth protein lied in 5’UTR, and the positive regulation in the course of skeletal muscle repair revealed a potential association with skeletal muscle cells development or gene expression of other growth related factors.
Key words: Pig, SRPK1, real-time PCR, inverse PCR (I-PCR), bioinformatics analysis, skeletal muscle damage model.