CDNA Cloning And Tissue Expression Characteristics Of BTG1Gene And TCAP Gene In Tianfu Goat

In this study, based on ovine BTG1gene mRNA sequences and bovine TCAP gene mRNA sequences, the first successfully cloned the BTG1gene and TCAP gene in Tianfu goat, the BTG1gene cDNA sequence of a length of593bp and624bp TCAP genecDNA sequence, the sequence was submitted to the NCBI, the two gene accession number (JQ247191.1, and JN129836.1).BTG1, which was593bp in length with an open reading frame(ORF)516bp long and encoded171amino acids, protein molecular weight19194.9Da, theoretical isoelectric point is8.35;cDNA sequences of TCAP was624bp in length with an open reading frame (ORF)504bp long and encoded167amino acids,protein molecular weight18970.5Da,theoretical isoelectric point of5.50.Bioinformatics analysis showed that:the Tianfu goat BTG1cDNA sequence and consistency of sheep, cattle, pig, rat and human are as follows:99%,99%,97%,95%,95%. Its amino acid and consistency of sheep, cattle, pig, rat and human, respectively:100%,100%,99%,98%,99%. The Tianfu goat TCAP cDNA sequence of cattle, swine and human consistency, respectively:97%,94%,91%. Amino acid and consistency of cattle, pig and human, respectively:97%,95%,93%; BTG1protein and TCAP protein amino acids are hydrophilic; BTG1protein contains a transmembrane domain, and contains two functional domains. TCAP protein does not contain transmembrane domains, contain a conserved domain; BTG1amino acid sequence containing four Ser phosphorylation sites, two Thr phosphorylation sites, two Tyr phosphorylation sites, five specific proteins kinase C phosphorylation sites; the TCAP amino acid sequence contains eight Ser phosphorylation sites, three Thr phosphorylation sites, a Tyr phosphorylation sites, three specific protein kinase C phosphorylation sites; BTG1amino acid sequence contains the a-helix:37.43%random coil:45.61%, extension of the fragment:16.96%; the TCAP amino acid sequence contains the a-helix:31.73%random coil:49.10%, extension of the fragment:19.16%.The expression of BTG1in patial selected tissues in Tianfu goat were analysed by real-time fluorescence quantitative PCR. results showed that BTG1were all expressed in eight selected tissues(heart, liver, spleen, lung, kidney, longissimus, leg muscle,abdominal muscle), when the Tianfu goat was of4-month age, the highest expression was observed in spleen,which was significantly higher than that other tissues (P<0.01),the differences of the liver, the kidney and between the lungs are not significant (P>0.05),they were significant higher than the myocardium, leg muscles, longissimus and abdominal muscle(P<0.01).the difference of the myocardium,longissimus, leg muscles and abdominal muscle is no significant (P>0.05), expression of BTG1gene is low in the muscle,the expression of BTG1decreased in the spleen with the rise of the age, the expression BTG1first rose then fell in the liver, the expression of BTG1rose the longissimus and leg muscle.The expression of TCAP in patial selected tissues in Tianfu goat were analysed by real-time fluorescence quantitative PCR. results showed that TCAP were all expressed in eight selected tissues(heart, liver, spleen, lung, kidney, longissimus, leg musele,abdominal muscle), express of TCAP gene was rich in the Muscle tissue when the Tianfu goat was4-month age,the highest expression was observed in longissimus,which was significantly higher than that of kidney, lung,spleen and liver(P<0.01),the differences of longissimus,leg muscle, and myocardium are not significant (P>0.05),the difference between myocardium and abdominal muscle is not significant (P>0.05),leg muscles, myocardial and abdominal significant higher than kidney, lung, spleen and liver,the difference of expression were no significant in the spleen and liver, lungs, liver and spleen.the expression of TCAP decreased in muscle with the rise of the age.