山羊DLK1基因分子克隆、生物信息学分析及表达研究

doi:10.11843/j.issn.0366-6964.2016.03.007

Abstract

Abstract:

The present study aimed to clone DLK1 gene of goat，predict the structure and function of peptone encoded by DLK1 gene，further investigate its expression regulation in organs or tissues of goat and analyze the correlation between DLK1 mRNA expression and intramuscular fat(IMF) content in muscles.Jianzhou Da’er goat was used as experimental animals and the RT-PCR technique was employed to amplify the DLK1 gene.The biological characteristics of this gene were analyzed by bioinformatics，and the tissue expression specificity as well as the development stage expression specificity of this gene were analyzed using the fluorescence quantitative PCR.The results showed that the DLK1 cDNA was 1 137 bp in length（GenBank accession No.：KP686197），927 bp for ORF and 308 amino acids for protein encoded.The nucleotide sequence and the deduced amino acids sequence of goat DLK1 shared 97% homology with the DLK1 of cattle.Accordingly，the close genetic relationship between goat and cattle was indicated by the phylogenetic tree analysis.The DLK1 protein was an unstable，acidity and hydrophobicity protein.There were 8 phosphorylation sites and 1 N-glycosylation site within the DLK1 protein.This protein was located in endoplasmic reticulum(44.4%)，golgi(33.3%) and plasma membrane(22.2%)，respectively through the sub-cellular level prediction.It was a secretory transmembrance protein.It had 5 domains of the EGF family，which had 2 EGF-CA of EGF family conservative domains.Moreover，the secondary structure of the DLK1 protein was composed of 87.01% random coil and 12.99% β-sheet，indicating an unconventional protein.The results of real-time PCR exhibited that the DLK1 gene was expressed in various tissues at different levels.The mRNA expression level of the DLK1 gene was highest in kidney and lowest in fat.The mRNA expression level of the DLK1 gene in longissimus dorsi of lamb was not significant higher than that in longissimus dorsi of youth and adult goat (P＞0.05).The results of correlation analysis showed that the mRNA expression level of the DLK1 gene in longissimus dorsi was correlated negatively with IMF content(R=-0.6，P＜0.05)，in leg was not correlated positively with IMF content(R=0.2，P＞0.05)，in arm triceps was not correlated negatively with IMF content(R=-0.2，P＞0.05).The results suggest that the DLK1 gene may play an important role in the IMF deposition in goat，which would lay a foundation for further studies about the function of DLK1 gene in IMF deposition.

CLC Number:

S827
S813.3

LIAO Hong-hai，LIN Ya-qiu，WU Yang-nan，ZHU Wu-zheng，LI Qian，WANG Yong，CHEN Juan. Molecular Cloning，Bioinformatics Analysis and Expression of DLK1 Gene in Goat[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(3): 467-476.

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Molecular Cloning，Bioinformatics Analysis and Expression of DLK1 Gene in Goat

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