PCR技术在麦哲伦企鹅性别鉴定上的应用

Application of PCR Technique in Sex Determination of Magellanic Penguins(Spheniscus magellanicus)

以青岛水族馆7只麦哲伦企鹅的新鲜血液为对象提取基因组DNA,根据基因组数据设计性别特异引物P2/P8,对其CHD基因进行PCR扩增,回收扩增的目的片段进行测序,获得这些麦哲伦企鹅的性别相关基因序列。将这7只麦哲伦企鹅的CHD基因序列与GenBank上公布的短趾雕的CHD基因序列进行同源性比对分析,建立遗传进化树,初步确定编号为1572、1573、1574、1575、36的麦哲伦企鹅鉴定为雄性,编号29和1577号麦哲伦企鹅为雌性。研究表明,雌雄麦哲伦企鹅CHD基因的内含子基因序列存在差异,P2/P8引物可扩增该保守序列,基于CHD基因建立的分子生物学方法可以较好地对麦哲伦企鹅实现性别的鉴定。

We studied the sex determination of Magellanic penguin(Spheniscus magellanicus)using seven birds at Qingdao Aquarium.We extracted genome DNA from fresh blood,and amplified the CHD gene using the PCR method with the primer-pair P2/P8 specially designed based on the genome sequences.We obtained the gender-related gene sequences of these Magellanic penguins after sequencing of the purified amplicons.We compared the homology between the CHD gene sequences of the 7 Magellanic penguins and the those of Circaetus gallicus published by GenBank,and constructed a genetic evolutionary tree based on the CHD gene sequences of the two species.We determined that Magellanic penguins#1572,#1573,#1574,#1575,and#36 were males,and#29 and#1577 were females.This study showed that there were differences in the intron sequence of the CHD gene between male and female Magellan penguins,and primer-pair P2/P8 could amplify the conserved sequences.The molecular biological method based on the CHD gene proved effective in determining the gender of Magellan penguins.