Xiang Wenjin, Shang Ying, Wang Qin, Xu Yuancong, Zhu Pengyu, Huang Kunlun, Xu Wentao
Beijing Laboratory of Food Quality and Safety, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China.
Yunnan Institute of Food Safety, Kunming University of Science and Technology, Yunnan 650500, China.
Food Chem. 2017 Nov 1;234:472-478. doi: 10.1016/j.foodchem.2017.05.038. Epub 2017 May 8.
The genes commonly used to determine meat species are mainly mitochondrial, but the copy numbers of such genes are high, meaning they cannot be accurately quantified. In this paper, for the first time, the chromosomal gene Actb was selected as an endogenous reference gene for chicken species. It was assayed in four different chicken varieties and 16 other species using both qualitative and quantitative PCR. No amplification of the Actb gene was found in species other than chicken and no allelic variations were detected in chicken. Southern blot and digital-PCR confirmed the Actb gene was present as a single copy in the chicken genome. The quantitative detection limit was 10pg of DNA, which is equivalent to eight copies. All experiments indicated that the Actb gene is a useful endogenous reference gene for chicken, and provides a convenient and accurate approach for detection of chicken in feed and food.
常用于确定肉类物种的基因主要是线粒体基因，但此类基因的拷贝数很高，这意味着无法对其进行准确量化。本文首次选择染色体基因Actb作为鸡物种的内参基因。使用定性和定量PCR在四个不同鸡品种和其他16个物种中对其进行了检测。在鸡以外的物种中未发现Actb基因的扩增，在鸡中也未检测到等位基因变异。Southern印迹和数字PCR证实Actb基因在鸡基因组中以单拷贝形式存在。定量检测限为10pg DNA，相当于八个拷贝。所有实验表明，Actb基因是一种用于鸡的有用内参基因，并为饲料和食品中鸡的检测提供了一种方便且准确的方法。
