中国地方家鸭ZW性染色体上CHD1基因序列差异的检测新方法

doi:10.11843/j.issn.0366-6964.2013.05.021

畜牧兽医学报 ›› 2013, Vol. 44 ›› Issue (5): 817-822.doi: 10.11843/j.issn.0366-6964.2013.05.021

中国地方家鸭ZW性染色体上CHD1基因序列差异的检测新方法

胡艳¹，徐文娟¹，宋迟¹，宋卫涛¹，单艳菊¹，陈文峰²，李慧芳^1*

（1. 江苏省家禽科学研究所，江苏省家禽遗传育种重点实验室，扬州 225003； 2. 江苏腾达源农牧有限公司，姜堰 225538）

收稿日期:2012-07-16 出版日期:2013-05-23 发布日期:2013-05-23
通讯作者: 李慧芳，博士，研究员，E-mail:lhfxf_002@yahoo.com.cn
作者简介:胡艳（1972-），女，湖南永州人，博士，高级兽医师，主要从事家禽遗传育种和营养生理研究，Tel：0514-85599177，E-mail：huyan0128@126.com
基金资助:
国家自然科学基金（31172194）；江苏省科技支撑计划（BE2011329）；现代农业品种创新项目（CX(11)1030）；扬州市项目（YZ2010048）

A New Detecting Method for the Sequence Difference of CHD1 Gene on the Z and W Sex Chromosomes in Chinese Native Domestic Duck Breeds

HU Yan¹, XU Wen-juan¹, SONG Chi¹, SONG Wei-tao¹, SHAN Yan-ju¹,CHEN Wen-feng², LI Hui-fang^1*

（1. Key Laboratory of Poultry Heredity & Breeding, Jiangsu Institute of Poultry Science，Yangzhou 225003, China； 2. Jiangsu Tengdayuan Co. Ltd, Jiangyan 225538, China）

Received:2012-07-16 Online:2013-05-23 Published:2013-05-23

摘要/Abstract

摘要：

旨在发掘一种能高效和快速检测鸭ZW性染色体上CHD1基因序列差异的方法，以解决鸭产业中胚胎期无损伤性别鉴定、胚胎期性别人工鉴定误判和新生期翻肛鉴别损伤等技术问题。本研究利用鸟类CHD1基因序列的DNA扩增片段长度多态性特点，成功获得1对能够在中国地方家鸭品种中广泛使用的性别鉴定引物HPF/HPR，对所获得的PCR产物进行了克隆、测序和比对，并通过实例对该性别分子鉴定方法的有效性和准确性进行了验证。结果显示，经2% 琼脂糖凝胶电泳就能清晰区分由引物HPF/HPR获得大小为495 bp的CHD1-Z和351 bp的CHD1-W PCR产物，并且雌性（ZW型）同时出现明显的双条带，而雄性（ZZ型）则出现单一条带。经证实，本研究提供的检测中国地方家鸭ZW性染色体上CHD1基因序列差异的方法直观可靠，同时也为中国地方家鸭的性别分子生物学鉴定提供了一个高效准确的分子遗传标记。

Abstract:

In the current study, a new method for detecting the sequence difference of duck Chromo-helicase-DNA-Binding 1 (CHD1) genes on Z and W sex chromosomes was established through amplifying DNA fragment length polymorphism, which aimed to solve some technical problems, such as the sex identification with nondamage, the misclassific rate of artificial sex identification during embryonic period and the injury coming from the rectal identification during neonatal period. In this study, an improved gender identification method was developed by DNA amplified fragment length ploymorphism between CHD1-Z and CHD1-W genes of birds. The gender identification PCR primers HPF/HPR were used, and the PCR product was cloned, sequenced and blasted; then the effectiveness and accuracy of this method had been explored by examples. 2% agarose gels were used to easily distinguish the 495 bp CHD-Z and the 351 bp CHD-W PCR amplicons, and the female (ZW) displayed two visible bands, whereas only a single band was found in the male (ZZ). The results indicate that the new detecting method for detecting the sequence difference of duck CHD1 genes is visual and reliable, and the molecular marker provided by this study is highly efficient and precise in the sex molecular biological identification of Chinese native domestic duck breeds.

中图分类号:

S831.2

胡艳，徐文娟，宋迟，宋卫涛，单艳菊，陈文峰，李慧芳. 中国地方家鸭ZW性染色体上CHD1基因序列差异的检测新方法[J]. 畜牧兽医学报, 2013, 44(5): 817-822.

HU Yan, XU Wen-juan, SONG Chi, SONG Wei-tao, SHAN Yan-ju,CHEN Wen-feng, LI Hui-fang. A New Detecting Method for the Sequence Difference of CHD1 Gene on the Z and W Sex Chromosomes in Chinese Native Domestic Duck Breeds[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2013, 44(5): 817-822.

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中国地方家鸭ZW性染色体上CHD1基因序列差异的检测新方法

A New Detecting Method for the Sequence Difference of CHD1 Gene on the Z and W Sex Chromosomes in Chinese Native Domestic Duck Breeds

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