藏鸡KLF15基因克隆、组织表达谱及其表达与肌内脂肪含量相关性的研究

doi:10.11843/j.issn.0366-6964.2019.02.004

摘要/Abstract

摘要：

旨在获得藏鸡KLF15基因序列，阐明其组织和时序表达谱，并分析该基因表达与肌内脂肪（IMF）含量的关系。本研究选取1、81、119、154、210日龄健康公母藏鸡各5只为试验动物，利用RT-PCR技术克隆KLF15基因，利用生物信息学软件分析基因生物学特征，利用实时荧光定量PCR（fluorescence quantitative PCR，qPCR）检测KLF15基因在藏鸡不同组织、不同发育阶段的表达谱，并分析其表达水平与肌内脂肪（intramuscular fat，IMF）含量的相关性。结果显示，克隆得到藏鸡KLF15基因序列长度为1 288 bp（GenBank登录号：KY747450），开放阅读框为1 212 bp，编码403个氨基酸，是具有3个锌指结构的亲水不稳定碱性蛋白质。藏鸡KLF15基因与原鸡的核苷酸和氨基酸的同源性均为99%。KLF15 mRNA在藏鸡各个组织中广泛表达，但在肺组织中表达水平最高，极显著高于其他组织（P<0.01）。KLF15 mRNA在1日龄藏鸡胸肌组织中的表达水平高于其他各日龄段，并随着日龄的增长呈下降趋势；在119日龄腿肌组织中的表达水平最高，且整体表达水平的变化都呈先下降后上升再下降的趋势。藏鸡公鸡154日龄前KLF15基因的表达水平与其对应的IMF含量呈正相关，154日龄以后则呈负相关；藏鸡母鸡在各个日龄段则主要表现为微弱的负相关。上述结果为进一步揭示KLF15基因在藏鸡IMF沉积中的作用提供重要数据。

Abstract:

The objective of this study was to obtain the sequence of KLF15 gene of Tibetan chicken,clarify its tissue and temporal expression profiles and to analyze the relationship between its expression and the intramuscular fat (IMF) content. Ten healthy Tibetan chicken(5 cock and 5 hen) on 1, 81, 119, 154 and 210 days old were selected, respectively as experimental animals. RT-PCR technology was used to clone KLF15 gene of Tibetan chicken, and bioinformatics softwares were used to analyze the biological characteristics of KLF15. The expression profiles of KLF15 gene in different tissues and different developmental stages was detected by fluorescence quantitative PCR, and its correlation with IMF content was analyzed. The result showed that the cloned Tibetan chicken KLF15 gene sequence was 1 288 bp in length (GenBank accession number:KY747450), an open reading frame of 1 212 bp, encoding 403 amino acids, and it was a hydrophilic unstable basic protein with 3 zinc finger structures. The homology between the nucleotide and amino acid sequences of KLF15 gene of Tibetan chicken and Gallus gallus were 99%. KLF15 mRNA was widely expressed in all detected tissues of Tibetan chicken, and the expression level was the highest in lung, which is significantly higher than that in other tissues (P<0.01). The expression level of KLF15 gene in breast muscle of Tibetan chicken of 1 day old was higher than that of the other age groups, and declined with the increasing age. The expression level of KLF15 gene in leg muscle of Tibetan chicken of 119 days old was higher than that of the other age groups, and the change of the overall expression level showed a trend of decreasing first, then rising, finally falling. The expression level of KLF15 gene before 154 days old was positively correlated with IMF content in Tibetan cocks,after 154 days old, the correlation was reverse, and the Tibetan hens mainly showed a weak negative correlation in all age groups. These results provide the basic data for further research on the role of KLF15 in IMF deposition of Tibetan chicken.

中图分类号:

S831.2

王英明, 徐亚欧, 王志敏, 徐珑洋, 杨磊, 林亚秋. 藏鸡KLF15基因克隆、组织表达谱及其表达与肌内脂肪含量相关性的研究[J]. 畜牧兽医学报, 2019, 50(2): 261-270.

WANG Yingming, XU Ya'ou, WANG Zhimin, XU Longyang, YANG Lei, LIN Yaqiu. Studies on the Cloning of KLF15 Gene, Tissue Expression Profile and the Association between Its Expression and Intramuscular Fat Content in Tibetan Chicken[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(2): 261-270.

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