CDNA Cloning And Tissue Expression Of TNNT1、TNNT2 And TNNI2 In Tianfu Goat

Skeletal muscle is the most important. component of animal body part, accounting for 40% of the body’s production of meat animals, but also one of the important factors affecting meat quality and animal muscle, so the impact of meat production in skeletal muscle gene is important genes. Troponin 12 type (TNNI2) gene and troponin T1 (TNNT1) is one of the important genes affect the performance of the animals slaughtered as fast skeletal muscle genes play a role in the regulation of skeletal muscle in animal growth and development process,and troponin T2 (TNNT2) as cardiac subtypes, has great research value in terms of heart disease.The results show:TNNI2 gene open reading frame (ORF) of 549 bp, encoding 187 Amino acid sequence has a conserved domain,there are six phosphorylation sites, protein secondary structure to random coil and α- helix,the sequence submitted to NCBI accession number obtained:KF939314; TNNT1 gene open reading frame of 759 bp, encoding 252 amino acids, the amino acid sequence has seven phosphorylation sites, a conserved domain of protein secondary structure has only random coil and α- helix, no extension fragments obtained submit NCBI accession number:KF939315,TNNT2 gene open reading frame of 858bp,encoding 285 amino acid sequence containing 15 phosphorylation sites,a conserved domain structure of the protein is mainly corpus and α- helical crimp random submitted NCBI accession number obtained KF939316.Tianfu goat TNNT1 on amino acid sequence alignment showed that cattle, who is a dolphin species, genus sheep, antelope species, eight Bandicota genus, genus panda, pig genus,Felis genus and the alpaca similarity respectively 94%,92%,93%, 72%,85%,91%,92%,93%,93% and 91%, and the evolutionary relationship TNNT1 genes in other species, Tianfu goat TNNT1 genetic relationship with the pig, followed by sheep, Tibetan antelope, eight tooth mouse and cattle, domestic cats, ferrets and pandas points in a primate clade; TNNT2 amino acid sequences were compared, the results show Tianfu goat and cattle are human genus, buffalo genus, gibbon genus, rhino genus canis, Panda is a pig is, ferrets and cats are the similarities are respectively 94%,88%,95%,88%,89.%,89%,89%.91%,88% and 89%; evolutionary relationships TNNT2 genetic relationship between Tianfu goat ruminant cattle recently, with humans, chimpanzees and gibbons and other primates together as one, then and ferrets and clustered into a panda, pig and minke whales clustered into one, clustered into a wolf and walrus; TNNI2 amino acid sequence and walrus, pig, rat, rabbit, mouse, human, guinea pigs, cattle the similarity of chimpanzees and dolphins were 97%,96%,96%,95%,94%,96%,94%,98%,92%and 92% amino acid sequence constructed TNNI2 phylogenetic tree, Tianfu goat and cattle are recently, followed walrus, human, pig, and dolphin furthest.Real-time quantitative RT-PCR analysis of the expression of three genes technical Tianfu goat in the part of the organization, the results showed that:TNNI2 Tianfu selected genes in goat 11 organizations,the highest expression of the abdominal muscles, followed by the diaphragm, and other organizations the difference was significant (P<0.01); in 1 day,183days,365days and 730days, butl2 months abdominal tissue of age with other significant difference (P<0.01). TNNT1 gene expression in the lung in the highest, low expression in the heart; in the four ages of 12-month-old lung expression levels than other age groups. TNNT2 gene is highly expressed in the heart, followed by the liver and lungs, the differences between the other eight organizations was not significant,in the heart, TNNT2 gene expression levels increased with age growth.Western blot (Westren Bloting) analysis in 365-day-old goat Tianfu eight organizations TNNT1 protein expression showed that, in the eight organizations were detected signal, which in addition to the diaphragm weak signals, other than the strong; only differences in the expression levels between the soleus muscle and significantly (P<0.05), the comparisons between the various other organizations showed significant difference (P<0.01).