The Pelican's Gender Identification Method And Its Genetic Clustering Analysis

Pelican is a second class nation-level pretected animal, which is monomorphic birds, is the male and female same appearance type. it is difficult to directly determine gender according to physical characteristics. So the phenomenon of mismatch had been happened in the breeding and breeding, reproduction rate of pelican is low in zoo.Conservative CHD gene to identify birds sex is that CHD has two homologous copies CHD-W and CHD-Z, the exon size of CHD-Z and CHD-W gene is close, but the intron length was different. So using PCR amplification reaction of introns part of its expansion to judge sexuality. If the PCR results showed a band, namely for CHD-Z, can be judged as a male, if shows two bands, for CHD-Z, CHD-W, namely can determine for females. The current widespread use of PCR primers P2/P8, 2550F/2550 R on the pelican sex determination is not specific, so the purpose of this study is to design specific primers of sex determination in the pelican, so that stable and reliable to identify the pelican gender,improve the breeding efficiency of pelicans.This experiment amplify genome of the pelican using primer 2550 F / 2718 R, then we can get the CHD gene sequences of pelican by recycling and sequencing, using Chromos software and DNAstar package and Clustal W, as well as in NCBI Blast with comparative analysis, design a specific primers for pelican. there are 13 primers designd for pelican in this experiment, three of them are for CHD-Z gene specific primers, 7 are for CHD-W gene specific primers, 3 are specific for CHD gene. Through to the pelican individuality other validation results show that, primer combination Z2/W3, Z3/W3 experimental accuracy is100%, 90.47%; Primer T2, T3 biological experimental accuracy is 95.23%, 100%. In addition, primers in DNA template concentration of 1 ng/?L, primer can still effective.Therefore, this study established research methods strong specificity, high accuracy, can be applied to the pelican sex determination.In the lab of the pelican, CHD-Z, CHD-W fragment sequence and chicken,Pycnonotus taivanus, crested ibis, vultures, vultures corresponding homologous sequence of common birds such as cluster analysis. Results show that the pelican segments of CHD-W and CHD-Z in the experiment genetic evolutionary relationships with other birds, and they have great similarity with the crested ibis.