Cloning, Sequence Analysis And Partial Tissue Expression Of CTSL,CTSH And CTSF Gene In Tianfu Goat

Meat quality traits are important economic traits on livestock production,so the gene affecting meat quality traits of livestock is becoming a hot research topic in recent years. Cathepsin family belongs to cysteine proteases family.It can not only hydrolyzes proteins in muscle fibers,but also makes animal connective tissue loose.Because of this,it can indirectly associated with tenderness. Some cathepsin family members have related reports in pigs, cattle and fish,but no report on goat. Previous research shows that cathepsin gene family members can be a candidate gene for meat quality traits in livestock.In this study, RT-PCR cloning and sequencing methods were combined to obtain the Tianfu goat CTSL gene, CTSH gene, CTSF gene and their whole coding sequence. The results show that Tianfu goat CTSL gene has a length of 1103bp Including a 1005bp coding region and encoding 334 amino acids, it was then deposited into the GenBank database （GenBank accession No. KF953544.1）. CTSL has signal peptide of amino acids, containing 17 phosphorylation sites, belonging a hydrophilic non-transmembrane proteins. Sequence prediction indicates that CTSL amino acid contains two conserved domains, namely inhibitor₁29 domain and Pept_Cl domain. Amino acid sequence homology analysis showed that it is a high similarity between Tianfu goat CTSL amino acid sequences and other mammals. Amino acid sequence homology analysis and phylogenetic analysis showed that Tianfu goat CTSL and sheep CTSL has close genetic relationship. Its secondary structure has a helix and go-curl as the main.The results show that Tianfu goat CTSH gene has a length of 1043bp Including a 1008bp coding region and encoding 335 amino acids, it was then deposited into the GenBank database （GenBank accession No.KJ650499）. CTSH has signal peptide of amino acids, containing 11 phosphorylation sites, belonging a hydrophilic non-transmembrane proteins. Sequence prediction indicates that CTSH amino acid contains two conserved domains, namely inhibitor₁29 domain and Pept_C1domain. Amino acid sequence homology analysis showed that it is a high similarity between Tianfu goat CTSH amino acid sequences and other mammals. Its secondary structure has a helix and go-curl as the main.The results show that Tianfu goat CTSF gene has a length of 1477bp Including a 1383bp coding region and encoding 460 amino acids, it was then deposited into the GenBank database （GenBank accession No.KJ650498）. CTSF has signal peptide of amino acids, containing 28 phosphorylation sites, belonging a hydrophilic non-transmembrane proteins. Sequence prediction indicates that CTSF amino acid also contains two conserved domains, namely inhibitor₁29 domain and Pept_C1 domain. Amino acid Amino acid sequence homology analysis and phylogenetic analysis showed that Tianfu goat CTSF and sheep CTSF has close genetic relationship. Its secondary structure has a helix and go-curl as the main.Real-time quantitative respectively analyzes the eight tissues relative expression of Tianfu goat CTSL gene, CTSH gene and CTSF gene at 12 months. It was also analyzed by using the same way that the 3 genes relative expression at ditterent time on lung tissue and crureus. The results showed that CTSL gene, CTSH gene and CTSF gene was all expressed in eight organizations. Their highest expression levels were tested in lung tissue. In lung tissue, the relative expression of all three genes increased at first and then decreased with age increasing. But in the leg muscle tissue, the CTSL genes and gene CTSH relative expression decreased at first and then rise with age increasing. The relative expression of the CTSF gene firstly increased and then reduced again rising.Tianfu goat CTSH eight organizations relative expression in 12-month-old was analysed by Western blotting. The results showed that all eight organizations have been detected expression. The relative expression is highest in crureus, significantly higher than other tissues （P<0.01）,then following the pectoral muscle, liver, kidney, abdominal muscle, diaphragm, myocardium and lungs from high to low.