藏山羊SFRS18基因克隆、表达及其与肌内脂肪含量相关性研究

doi:10.16431/j.cnki.1671-7236.2015.11.029

Abstract

Abstract: In the present study,SFRS18 gene CDS region of Tibetan goat was cloned and analyzed by bioinformatics method,further its expression regulation in organs or tissues of goat were investigated and the correlation between SFRS18 mRNA expression and intramuscular fat (IMF) content in muscles was analyzed.Thereby providing data for further investigation on the gene functions in goat IMF deposition.The RT-PCR technique was employed to amplify the SFRS18 gene of Tibetan goat.Physicochemical property,structure and homology of SFRS18 encoded protein were analyzed by bioinformatic methods,and the tissue expression specificity as well as the development stage expression specificity of this gene were analyzed using the Real-time fluorescence quantitative PCR.The results showed that 1299 bp for SFRS18 cDNA,1272 bp for ORF and 423 amino acids for protein encoded.The formula of SFRS18 protein was C₂₀₀₃H₃₄₂₄N₇₆₀O₆₉₆S₄,and the molecular weight of SFRS18 protein was 49.42 ku,and its theoretical isoelectric point was 11.20.SFRS18 protein was unstable and hydrophilic,without signal peptide.There were 103 phosphorylation sites,2 N-glycosylation sites and 39 O-glycosylation sites within the SFRS18 protein.This protein respectively located in nuclear (82.6%),cytoplasmic (8.7%),cytoskeletal (4.3%) and plasma membrane (4.3%) through the sub-cellular level prediction.It was an untransmembrance protein.Moreover,the secondary structure of the SPRS18 protein was composed of 99.29% random coil and 0.71% α-helix,indicating an unconventional protein.The nucleotide sequence and the deduced amino acids sequence of Tibetan goat SFRS18 shared 99% homology with the SFRS18 mRNA of Bos taurus,Ovis aries and Bubalus bubalis.Accordingly,the close genetic relationship between Tibetan goat and Bos taurus was indicated by the phylogenetic tree analysis.The results of Real-time PCR exhibited that the SFRS18 gene was expressed in various tissues at different levels.The mRNA expression level of the SFRS18 gene was the highest in spleen and the lowest in longissimus dorsi.The results of correlation analysis showed that the mRNA expression level of the SFRS18 gene in longissimus doris and in crureus was correlated positively with IMF content (r=0.081,P<0.05;r=0.373,P<0.05).The SFRS18 gene was involved in the regulation of IMF deposition in goat.

Key words: Tibetan goat; SFRS18 gene; clone; intramuscular fat (IMF)

CLC Number:

LIAO Hong-hai, LIN Ya-qiu, LI Qian, WU Yang-nan, WANG Yong. Cloning,Expression Analysis of SFRS18 and Correlation Analysis between its mRNA Expression and Intramuscular Fat Content in Tibetan Goat[J]. , 2015, 42(11): 3016-3025.

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Cloning,Expression Analysis of SFRS18 and Correlation Analysis between its mRNA Expression and Intramuscular Fat Content in Tibetan Goat

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