鸡ev21占位区和未占位区重复序列结构与羽速表型关系研究

doi:10.11843/j.issn.0366-6964.2017.06.003

畜牧兽医学报 ›› 2017, Vol. 48 ›› Issue (6): 990-999.doi: 10.11843/j.issn.0366-6964.2017.06.003

鸡ev21占位区和未占位区重复序列结构与羽速表型关系研究

张乐超¹, 王晗³, 张秀玲^1,4, 刘春杨¹, 王麒¹, 周荣艳¹, 李祥龙^2*, 李兰会^1*

1. 河北农业大学动物科技学院, 保定 071001;
2. 河北科技师范学院, 秦皇岛 066600;
3. 西北农林科技大学动物科技学院, 杨凌 712100;
4. 中国农业科学院北京畜牧兽医研究所, 北京 100193

收稿日期:2017-01-06 出版日期:2017-06-23 发布日期:2017-06-23
通讯作者: 李兰会,副教授,硕士生导师,主要从事动物遗传育种研究,E-mail:lanhuili13@163.com;李祥龙,教授,博士生导师,主要从事动物遗传育种研究,E-mail:lixianglongcn@yahoo.com
作者简介:张乐超(1991-),男,河北藁城人,硕士,主要从事动物遗传育种研究,E-mail:1596653383@qq.com
基金资助:
河北省现代农业技术体系蛋鸡产业创新团队（HBCT2013090206）；河北省自然科学基金（C2015204176）

Relation between Feather Speed and Gene Structure of Repeated Sequences of ev21 Occupied-site and Unoccupied-site Regions in Chicken

ZHANG Le-chao¹, WANG Han³, ZHANG Xiu-ling^1,4, LIU Chun-yang¹, WANG Qi¹, ZHOU Rong-yan¹, LI Xiang-long^2*, LI Lan-hui^1*

1. College of Animal Science and Technology, Hebei Agricultural University, Baoding 071001, China;
2. Hebei Normal University of Science & Technology, Qinhuangdao 066600, China;
3. College of Animal Science and Technology, Northwest A&F University, Yangling 712100, China;
4. Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China

Received:2017-01-06 Online:2017-06-23 Published:2017-06-23

摘要/Abstract

摘要：

本试验旨在探讨鸡羽速基因型Hae Ⅲ酶切鉴定方法的分子基础。以羽型明确的6个品系（太行鸡、坝上长尾鸡、大午粉鸡、海兰灰鸡、海兰褐鸡、海兰灰祖代四系鸡）313份鸡基因组为模板，利用Hae Ⅲ酶切的RFLP试验进行羽速验证，并对ev21占位区（OS）和非占位区（US）共有序列以及OS区特有序列进行酶切试验。结果表明：1）Blast分析发现1 450 bp为鸡ev21的US区域片段。海兰灰及其祖代四系和大午粉祖代的羽速基因型与Hae Ⅲ酶切结果完全一致，但太行慢羽公鸡、慢羽母鸡、坝上长尾慢羽公鸡和海兰褐慢羽鸡的一致率分别为40.0%、27.6%、28.6%和0.0%；2）太行鸡和坝上长尾鸡ev21的OS和US区共有序列538 bp酶切鉴定结果与表型一致率达到92%以上，其他品系（除海兰褐快羽公鸡为0.0%）均为100.0%；3）鸡ev21的OS区特异序列1 440 bp的扩增阳性率在太行慢羽公鸡、慢羽母鸡和快羽公鸡中的阳性率分别为94.1%、65.5%和0.0%，在海兰灰祖代鸡中分别为100.0%和0.0%，并且1 440片段不能被Hae Ⅲ切开。综合分析6个品系ev21的OS和US区结构特征，认为US区Hae Ⅲ酶切位点变异不能作为慢羽和内源病毒ev21的鉴定依据，而OS区的ev21插入与1 440 bp序列Hae Ⅲ酶切位点的A→G突变和八碱基重复是紧密连锁的。

Abstract:

The aim of this study was to explore the molecular basis of Hae Ⅲ digestion detection of genotypes of chicken feather speed in chicken. Three hundred and thirteen individuals in 6 strains of chickens (Taihang, Bashang Long-tail, Dawufen, Hy-line Gray, Hy-line Brown, progenitor of Hy-line Gray) with definite phenotype of feather speed were detected with RFLP. The common region of occupied site(OS) and unoccupied site(US) for 538 bp, also the specific region of OS for 1 440 bp, were digested with Hae Ⅲ. The results showed that: 1) The fragment for 1 450 bp was the unoccupied-site repeated sequence for ev21 ascertained by Blast. The digestion detection results of Hae Ⅲ for 1 450 bp were completely consistent with the phenotype of HY-line Gray and progenitors of HY-line Gray and Dawufen, but their consistency with Taihang slow-feather cock, Taihang slow-feather hen, Bashang Long-tail slow-feather cock and HY-line Brown slow-feather chicken were 40.0%, 27.6%, 28.6% and 0.0%, respectively. 2) The consistency was above 92% between digestion results of 538 bp and phenotype in Taihang and Bashang Long-tail chicken, and the other strains were 100.0% except for HY-line Brown fast-feather cock with 0.0%. 3) The positive rate of 1 440 bp fragment PCR in slow-feather Taihang cock, Taihang hen and fast-feather Taihang cock was 94.1%, 65.5% and 0.0%, respectively, and it was 100.0% and 0.0% in slow-feather and fast-feather progenitors of Hy-line gray, respectively. The 1 440 bp fragment could not be digested by Hae Ⅲ and it could be a detection basis for ev21.The structure was analyzed comprehensively for the OS and US of ev21 in 6 strains of chicken and it was concluded that the site recognized by Hae Ⅲ enzyme in US could not be a detection base for slow-feather and ev21, while ev21 insertion in OS and the site for Hae Ⅲ digestion in 1 440 bp were linked with the mutation of A → G and 8 bases repeated.

中图分类号:

S831.2

张乐超, 王晗, 张秀玲, 刘春杨, 王麒, 周荣艳, 李祥龙, 李兰会. 鸡ev21占位区和未占位区重复序列结构与羽速表型关系研究[J]. 畜牧兽医学报, 2017, 48(6): 990-999.

ZHANG Le-chao, WANG Han, ZHANG Xiu-ling, LIU Chun-yang, WANG Qi, ZHOU Rong-yan, LI Xiang-long, LI Lan-hui. Relation between Feather Speed and Gene Structure of Repeated Sequences of ev21 Occupied-site and Unoccupied-site Regions in Chicken[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(6): 990-999.

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鸡ev21占位区和未占位区重复序列结构与羽速表型关系研究

Relation between Feather Speed and Gene Structure of Repeated Sequences of ev21 Occupied-site and Unoccupied-site Regions in Chicken

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