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| cDNA Cloning, Expression and Functional Analysis of Adiponectin Receptor 1 Gene (AdipoR1) in Xinong Saanen Dairy Goat (Capra hircus) |
| ,Jun LUO, , |
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Abstract Adiponectin can improve insulin sensitivity, regulate fatty acid metabolism and participate in the cell proliferation and differentiation. Based on the highly conservative region of adiponectin receptor 1 gene (AdipoR1) sequences from bovine (Bos taurus), human (Homo sapiens) and mouse (Mus musculus) in GenBank, the specific primers were designed. A full-length 2 032 bp cDNA sequence of AdipoR1 gene(GenBank accession GQ918145) was isolated and cloned in mammary gland of Xinong Saanen dairy goats (Capra hircus) using RT-PCR and RACE. AdipoR1 gene contained 185 bp 5' untranslated regions(UTR), 719 bp 3' UTR, and 1 128 bp coding sequences (CDS) which encode 375 amino acid (AA). AA sequence alignment showed that the AA of goat AdipoR1 had higher similarity (＞95%) with bovine, pig(Sus scrofa), mouse and human. The protein structure analysis showed that the predicted molecular weight was 42.44 kD, the isoelectric point was 7.19, and 7 transmembrane domains and no signal peptide were included in the entire sequence. Furthermore, expression analysis showed that AdipoR1 gene had the most abundant expression in lung, followed by the small intestine and the minimal expression was observed in heart. However, the expression of AdipoR1 in remaining eight tissues changed more gently. Expression analysis in the two different stages of mammary tissue revealed that there was significantly different (P＜0.05) between peak lactation and dry period. Goat mammary epithelial cells were treated with different concentrations of insulin and prolactin, and the result showed that the mRNA level of AdipoR1 in group treated with insulin decreased and there was the most obvious effect at insulin concentration of 50 nmol/L (P＜0.01); the mRNA level of AdipoR1 in group treated with prolactin increased and there was the most obvious effect at prolactin concentration of 100 mg/mL (P＜0.05). These results indicated that AdipoR1 had certain regulative effects in goat mammary epithelial cells. This work provides basic information to further reveal the function of AdipoR1 in goat mammary gland.
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Received: 23 May 2014
Published: 06 January 2015
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Corresponding Authors:
Jun LUO
E-mail: luojun@nwsuaf.edu.cn
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