The Study Of Related Genes Of Causing Abnormal Sexual Differentiation In The Chicken And Quail Intergeneric Hybrid

The case of chicken and quail is a kind of hybrid between a typical distant hybridization, and its hybrid sterility is an important manifestation of the hybrid embryo hatching early death, the death of concentrated time for 3-5 days, with the sexual differentiation period of overlap, in-depth study has also been confirmed that estrogen receptor (ER) in early embryonic death of hybrids play an important role. The key functions of the ER can control the expression of estrogen, estrogen in poultry to play a permanent role in sex differentiation, which can infer the effects of estrogen on the chicken and quail hybrids between the death of early embryonic plays an important role. Poultry into the body need estrogen hydroxy steroid dehydrogenase and 17a-hydroxylase the role of these genes in hybrid embryos hatching mechanism and the expression of early embryonic death of the relationship between early, can make the case for the chicken and quail hybrids between early embryonic death of a deeper understanding of the molecular mechanism of step. In this study on the base of Cloning of quail and 3p-HSD hybrid full-length sequence and P-450c17 gene fragment sequences, then carry out the relevant gene Quantitative on the positioning of these two genes in chickens, quails and hybrids of early embryonic development.The expression of results are as follows:1. Clone and sequence of 3β-HSD full-length gene and P-450c17 partial gene in the quails and their intergeneric hybrids of chicken.The genes of 3β-HSD and P-450c17:full-length sequence of chicken have been reported, some reports of quail sequence, no reports on the hybrid. This experiment is a hybrid between the 3β-HSD cds full-length sequence (submitted to GENBANK, ID as:FJ607243), quail 3β-HSD cds full-length sequence (submitted to GENBANK, ID as:FJ607242); intergeneric hybrid species of P-450c17 cds part of sequence (submitted to GENBANK, ID as:FJ607243), part of P-450c17cds quail sequence (GENBANK ID for submission:FJ607242). In this study, chicken, quail and hybrid embryos for research, the use of extracted total RNA, RT-PCR technology,3β-HSD and P-450c17 gene sequence analysis of cloning, protein prediction.The results showed that compared with its hybrid, chicken had higher homology, while quail had lower homology compared with their hybrids. From these two proteins gene prediction results that they seemed be a globular protein. It's a greater similarity in the 3β-HSD protein between chicken and hybrid more than quail and their hybrid; while expressed similarity in quail and quail hybrids in P-450c17 protein, with a larger differentiation in the chicken and the other two.2. The positioning of early embryos 3β-HSD and P-450c17 in chicken and quail and their intergeneric hybrids3β-HSD and P-450c17 as the ER's one of the important regulatory enzyme, the expression of its specific site in the chicken and quail hybrids have been reported on. This experiment to chicken, quail and hybrid embryos for experimental materials, the adoption of the overall in situ hybridization method, from 3.5 to 5 days in chickens, quails and hybrids of early embryos and 3β-HSD gene expression in P-450c17 locate the site. The results showed that:3β-HSD and P-450c17 gene expression in the gonadal primordium surrounding reproductive ridge.3β-HSD gene in female hybrids 96h,102h and 108h embryos have the presence of blue-positive signal, that is the expression.3β-HSD expression in female embryos strong signal, while the male embryos in the 66h-168h also have expression; P-450c17 gene in female hybrids 96h,102h and 108h embryos exist positive blue signal, also is the expression. The results can be seen at the same time 3β-HSD in 72h,84h and 96h of female embryonic heart may also be part of the expression.3. Real Time PCR detection of ER,3β-HSD and P-450c17 three differential gene in chickens, quail and their intergeneric hybrids on the expression of the law.This report research the ER,3β-HSD and P-450c17 in the early embryonic expression, for the future of the biological function of these genes in-depth studies to determine the cause of death of early embryos to lay the foundation. In this experiment the five embryos were collected at 66h to 168h time of chickens, quail and their intergeneric hybrids, using the real-time PCR method to obtain the expression of target gene product line plans.The results showed that:1) ER gene:the vicinity of the peak period for expression in 108h chickens; the expression of the vicinity of the peak before the expression of instability in the 96h to 102h in quail; the expression of a lower peak in the vicinity of 72h hybrids, and then dropped to 96h when the expression of a higher peak. From the amount of expression, the chickens, quails and hybrids of the ER seems to be in the early embryonic expression of the existence of two peaks. The expression of chicken and quail is the higher peak the front than after the high. 2)P-450cl7 gene:108h chickens in the vicinity of the peak period for expression; quails express near the 96h peak; 90h hybrids reached near the peak of expression. The hybrid gene in the expression is the high, and the relatively low quail.3) 3β-HSD gene:chickens in the vicinity of 108h to reach the peak of expression; 120h quail in the vicinity of the peak period for expression; overall expression levels of hybrid low, 90h or less can be seen in the vicinity of a peak of expression, before and after changes in the expression of differences not obvious. Chicken, quail and hybrid gene expression of three peaks are in the 90h-108h. Hybrids showed a more pro-expression of this early trend; ER than the 3β-HSD, P-450c17 gene expression in advance, the amount of expression and their parents showed a greater difference in expression at the same time its time to its embryonic basically consistent with the time of death.