基于CHD基因序列分析的帝企鹅性别鉴定研究

Study on Sex Identification of Emperor Penguin(Aptenodytes forsteri) Based on Sequence Analysis of the CHD Gene

[目的]分析帝企鹅(Aptenodytes forsteri)CHD基因序列,以鉴定性别。[方法]采用苯酚∶氯仿抽提法提取6只未知性别的帝企鹅血液中的基因组DNA,运用P2/P8引物扩增CHD基因片段,将PCR产物克隆到T-Vector,利用NCBI的Blast程序,以短趾鹰(Circaetus gal-licus)同源序列为比对参照,将帝企鹅CHD基因片段序列与GenBank中的基因片段序列进行同源性比较分析,鉴定帝企鹅的性别。[结果]测序结果表明,CHD-Z和CHD-W基因的PCR产物大小分别为378、387 bp,雌雄结果并不明显,不易区分开来。Blast比对结果表明,帝企鹅LD、EK、ZF为雄性,帝企鹅DG、YA、YY为雌性。[结论]结合PCR扩增和测序分析CHD基因,是单态性鸟类性别鉴定的有效和准确的方法。

[Objective] The research aimed to analyze the CHD gene sequence of Emperor Penguin （Aptenodytesforsteri） to identify its sex. [ Method] The genetic DNA was extracted from the blood of 6 A.forsteri which sex was unknown. The PCR amplified fragments with primers P2/P8 were cloned in T-Vector. Taking homologous sequence of Circaetus gallicus as reference, the CHD gene sequence was compared to the homologues in GenBank to identify its sex. [ Result]The sequencing results showed that the PCR products of CHD - Z and CHD - W were 378,387 bp respectively, and sex character was not significant for dividing sex. The Blast comparative results showed that A.forsteri LD, EK and ZF was male, and A. forsteri DG , YA and YY was female. [ Conclusion]The method of combining PCR amplification and sequencing to analyze the CHD gene is an accurate and reliable way to determine sex of monomorphic avian.