Determination And Expression In C2C12Cells Of The CDS Of Porcine SkMLCK Gene

ObjectiveThe purpose is to determinate the CDS of porcine skMLCK gene and tobuild the pig skMLCK gene eukaryotic expression vector. The objective is toprobe the correlation between the expressions of porcine skMLCK gene and thedevelopment of muscle and to obtain the relationship between it and the traits ofpork preliminarily.MethodsBy referencing the predicted information on the NCBI and using the biologysoftware, the CDS is divided into two segments to undergo PCR amplification.By cloning and sequencing the porcine skMLCK gene CDS and splicing andsynthesizing the sequences, the porcine skMLCK gene eukaryotic expressionvectors are built. By liposome transfixing and screening, the stability expressskMLCK cells are acquired. The expressions of related gene are got byReal-Time PCR.ResultsThe pig skMLCK gene full-length CDS is obtained. Its length is1851bp.Thepig skMLCK gene eukaryotic expression vector is built successfully. It ispEGFP—N1—skMLCK. The stability express the gene skMLCK. cells areacquired.MHC2B gene is up regulated apparently. CFL2b and MEF2C are downregulated obviously. The mRNA expression correlation of the geneskMLCK-MEF2C and skMLCK-CFL2b are significant negative. The mRNAexpression level correlation of MEF2C-MHC2B is very significant positive.ConclusionsThe pig skMLCK gene expression significantly influenced the expressionof MHC2B, CFL2b and MEF2C gene. The research indicated the porcine pig skMLCK gene correlated to muscle fiber trait. This research can lay the basis forfurther study on the effect of the pig skMLCK gene in muscle fiber growth. A newdirection is proposed to improve and evaluate pork quality in gene level.