Analysis Dosage Compensation In Sika(Cervus Nippon) And Wapiti(Cervus Elaphus)-hybrid Female Calves On X Chromosome

Dosage compensation is for the male and female X chromosome which was the key to the balance. In orderto hybrid female offspring of X chromosome expression between sika and wapiti, we studied the high-qualityextraction of total RNA from the blood, using NCBI web on bovine or sheep Bruton’s tyrosine kinase (BTK),glucose-6-phosphate dehydrogenase (G6PD) and phosphoglycerate kinase (PGK1) gene as reference gene,designed primers to PCR about sika, wapiti and hybrid female calves of RNA by RT-PCR were synthesizedthrough reverse transcription cDNA. Get the part of gene sequence to analysis. This paper intends to investigatethe influence of bioinformatics analysis method on a hybrid female calves of sika and wapiti dosagecompensation. The results are as follows:1. Using peripheral venous blood extracted RNA, through different anticoagulants and pretreated methods tocompared. In the context of the Trizol method to extracted, Though the technology, electrophoresis, EB staining,and determining concentration the total RNA quality, Conclusion the best results after heparin anticoagulant bloodand red blood cell lysis treatmented, It can be used bioinformatics analysis and provided high quality materials onSika, Wapiti and hybrid female calves research.2. Using NCBI web, Get the bovine PGK1gene sequence. Designed primers to select sika, Wapiti and hybridfemale calves of RNA reverse transcription cDNA, which finally got into the sika deer, and hybrid calves PGK1gene length1572bp. Plus the above one were analyzed by gel detection, peak alignment and conserved sequenceprediction. The determination about PGK1gene is highly conservatism, and PGK encoding amino acid sequenceslength387aa. The expression of PGK1allelic genes were found in the red deer and hybrid female calves,Discussed the gene was escaped after X-choromosome inactivation. The gene only can be used as a referenceRNA quality.3. Through the analysis method combined with PGK1gene----the usage of sika deer, and after the hybridfemale calves BTK and G6PD gene amplification primer generation analysis, the length of the BTK gene wasamplified up to945bp, and the G6PD gene totals931bp. Predicted BTK gene primers could amplify the conservedScr kinase region, the amino acid sequences of length217aa; The amplification of G6PD gene contains two superfamily of proteins: G6PD_C and G6PD_N, the amino acid length were276aa and25aa. The hybrid female calvesof BTK and G6PD gene would happened dosage compensation effect, The effect was caused by X-choromosomeinactivation of Sika deer.