太行鸡FST基因克隆、序列特征及表达分析

doi:10.11843/j.issn.0366-6964.2021.011.008

畜牧兽医学报 ›› 2021, Vol. 52 ›› Issue (11): 3064-3075.doi: 10.11843/j.issn.0366-6964.2021.011.008

太行鸡FST基因克隆、序列特征及表达分析

张贝贝¹, 李梦霄¹, 马腾壑^1,2*, 李雪男¹, 魏佳荣¹, 康国磊¹, 王红娜¹, 刘超³, 王斌¹, 孙研研^4*

1. 河北工程大学生命科学与食品工程学院, 邯郸 056038;
2. 河北工程大学医学院, 邯郸 056038;
3. 河北天凯食品有限责任公司, 沙河 054100;
4. 中国农业科学院北京畜牧兽医研究所农业部动物遗传育种与繁殖(家禽)重点实验室, 北京 100193

收稿日期:2021-02-22 出版日期:2021-11-23 发布日期:2021-11-24
通讯作者: 马腾壑,主要从事家禽繁殖与分子育种研究,E-mail:matenghe@126.com;孙研研,主要从事鸡遗传育种研究,E-mail:yanyansun2014@163.com
作者简介:张贝贝(1996-),女,河北邢台人,硕士生,主要从事鸡遗传育种研究,E-mail:1051324922@qq.com
基金资助:
河北省蛋肉鸡农业产业技术体系（2018YFD0501300）；河北省科技厅项目（20326343D）；河北省自然科学基金（C2020402003）；河北省人社厅引进留学人员资助项目（C20190504）；河北省高等学校科学技术研究项目（ZD2018071）；邯郸市家禽工程技术创新中心（20312904034）

Cloning, Bioinformatics and Expression Analysis of FST Gene in Taihang Chickens

ZHANG Beibei¹, LI Mengxiao¹, MA Tenghe^1,2*, LI Xuenan¹, WEI Jiarong¹, KANG Guolei¹, WANG Hongna¹, LIU Chao³, WANG Bin¹, SUN Yanyan^4*

1. School of Life Sciences and Food Engineering, Hebei University of Engineering, Handan 056038, China;
2. Medical College, Hebei University of Engineering, Handan 056038, China;
3. Hebei Tiankai Food Company of Limited Liability, Shahe 054100, China;
4. Key Laboratory of Animal(Poultry) Genetics, Breeding and Reproduction of Ministry of Agriculture, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China

Received:2021-02-22 Online:2021-11-23 Published:2021-11-24

摘要/Abstract

摘要： 旨在基于生物信息学方法分析太行鸡卵泡抑素基因（follistatin，FST），并研究其在不同组织及卵泡不同发育时期表达差异，解析FST对太行鸡产蛋性能的影响。本研究使用RT-PCR对43周龄健康太行鸡FST基因编码区序列进行克隆和测序，运用生物信息学软件对其功能结构进行预测，荧光定量PCR技术对43周龄健康麻羽太行鸡FST在不同组织及不同发育时期卵泡中的表达差异进行分析，每组设3个重复，进行3次平行试验。结果显示，太行鸡FST基因编码区为1 032 bp，编码343个氨基酸，太行鸡FST蛋白与日原鸡同源性最高（100.0%），存在Sec信号肽，成熟蛋白具有FOLN和KAZAL_FS 2个保守结构域。该蛋白为亲水蛋白，蛋白分子式为C₁₆₁₉H₂₅₆₇N₄₅₉O₅₂₀S₄₄，分子量为38.192 6 ku，理论等电点为5.59，其中含量最高的是Cys （C），为10.50%；蛋白质二级结构含有α-螺旋、延伸链、β转角、无规卷曲，占比分别为16.33%、16.62%、5.25%、61.81%。FST基因在不同组织中均有表达，在肝中表达量最高。FST基因在各级卵泡表达量存在差异，其中，在大白卵泡中表达量最高（P<0.01），在卵泡选择前后表达量存在1.6倍表达差异。上述结果为研究太行鸡FST基因结构和功能提供了重要的基础数据，也为进一步挖掘太行鸡产蛋性能候选高产基因提供了参考。

关键词: 太行鸡, 卵泡抑素, 克隆, 表达, 生物信息学

Abstract: Follistatin (FST) gene of Taihang chicken was analyzed by bioinformatics methods and its expression differences in different tissues and follicles at different developmental stages was explored, and the effect of FST on the laying performance of Taihang chickens was analyzed. Coding region of FST gene was cloned and sequenced for 43-week-old healthy Taihang chicken, functional structure was predicted by bioinformatics softwares. Quantitative PCR (Q-PCR) was used to detect the expression differences of FST gene in different tissues and follicles at different developmental stages of Taihang chickens. Three replicates were set in each group and three parallel experiments were carried out. The results showed that the coding region of FST gene of Taihang chicken was 1 032 bp, encoding 343 amino acids. Homology analysis indicated that FST protein had the highest homology with Gallus gallus (100.0%), and both had Sec signal peptide. The mature protein had two conserved domains: FOLN and KAZAL_FS. Physicochemical properties analysis of FST protein showed that it was hydrophilic protein, whose molecular formula was C₁₆₁₉H₂₅₆₇N₄₅₉O₅₂₀S₄₄, molecular weight was 38.192 6 ku, and theoretical isoelectric point was 5.59, the highest content was Cys (C, 10.50%). The secondary structure consisted of α-helix, extended chain, β-corner and random coil, accounting for 16.33%, 16.62%, 5.25% and 61.81%, respectively. qPCR revealed that FST gene was expressed in all tissues, the highest expression was found in liver. During follicle development, FST gene was expressed highest in the large white follicles (P<0.01), there was a 1.6-fold difference in expression before and after follicle selection. These results provided important basic data for studying FST gene in Taihang chicken, and provided a reference for the further mining candidate gene for egg-laying performance of hens.

Key words: Taihang chickens, follistatin, cloning, expression, bioinformatics

中图分类号:

S831.2

张贝贝, 李梦霄, 马腾壑, 李雪男, 魏佳荣, 康国磊, 王红娜, 刘超, 王斌, 孙研研. 太行鸡FST基因克隆、序列特征及表达分析[J]. 畜牧兽医学报, 2021, 52(11): 3064-3075.

ZHANG Beibei, LI Mengxiao, MA Tenghe, LI Xuenan, WEI Jiarong, KANG Guolei, WANG Hongna, LIU Chao, WANG Bin, SUN Yanyan. Cloning, Bioinformatics and Expression Analysis of FST Gene in Taihang Chickens[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(11): 3064-3075.

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太行鸡FST基因克隆、序列特征及表达分析

Cloning, Bioinformatics and Expression Analysis of FST Gene in Taihang Chickens

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