关键词: 克隆, 表达, TNNC1, 大熊猫, 亚洲黑熊

Abstract: Troponin C type 1 (TNNC1)regulates the contraction of slow-twitch skeletal muscle and cardiac muscle,which may lead to differences in animal muscle growth,evolution,and function. In this study,the genomic sequences and cDNA of the TNNC1 gene of the giant panda (Ailuropoda melanoleuca)and the black bear (Ursus thibetanus)were cloned successfully using RT-PCR. The TNNC1 gene length of cDNA fragment of the giant panda contains an open reading frame of 486 bp encoding 161 amino acids and the length of the genomic sequence is 2 831 bp,contains six exons and five introns. Cloned TNNC1 gene cDNA fragment of the black bear contains a 486 bp open reading frame encoding 161 amino acids,the gene structure is 2 758 bp,also contains six exons and five introns. Alignment analysis indicated that the nucleotide sequences and the deduced amino acid sequences for TNNC1 gene of the two species are highly conserved with other 13 species which have been reported. Topology prediction shows that there are 5 Casein kinaseⅡphosphorylation sites,1 Protein kinase C phosphorylation site,1 N-myristoylation site,3 EF-hand calcium-binding domains and 1 N-glycosylation site in the TNNC1 protein of
the giant panda and the black bear. The cDNA of TNNC1 for the giant panda and the black bear were transfected into E. coli, the TNNC1 fused with the N-terminally His-tagged protein gave rise to the accumulation of an expected 23.5 kDa polypeptide, which was consistent with the predicted protein. The results of this study provide information to further explore the structure, function and evolution of TNNC1 gene and protein of the giant panda and the black bear.