Study Of Biological Function On A Novel Gene, GMPR2of Liaoning New-breeding Cashmere Goat And K26Antibody Preparation

Liaoning new breeding cashmere goat is our unique genetic resources, and it has a veryimportant economic value of production practices. Therefore, it is particularly important thatresearch the gene function related to liaoning new breeding cashmere goats economic traits.This is the first study of GMPR2gene in Liaoning new-breeding cashmere goat, aimingto study the function in the skin and the relationship with the cashmere fiber growth. Firstlybioinformatics analysis was used on this gene, and then positioning analysis was got by usingDIG-labeling in situ hybridization technique, and the relative quantitative analysis was carriedout through the real-time quantitative PCR technology. The expression of GMPR2gene inother organs was detected by RT-PCR technology. Finally gene expression is affected bydifferent concentration melatonin treatment in different time. So that GMPR2may indirectlycontrol the primary and secondry follicles stunted growth. In addition, the wool fiberformation of Liaoning new breeding cashmere goat is related to the expression of hair keratins(KIFS K). To study the protein of K26gene expression levels and expression position, thisexperiment build BL21-pET32a-K26expression vector. We extracted the recombinantprotein, and injected to New Zealand male rabbits for producing rabbit anti-K26geneantibody. This antibody can be as a follow-up study of the K26gene function tool. Importantresults are as follows:(1)The gene was really GMPR2gene though ORF Finder and Blast program, whichshowed highest homology with Bos taurus GMPR2gene.(2) It showed that the signals of GMPR2gene expressed in inner root sheath of primaryand secondary follicles in situ hybridization analyses of follicles.(3)GMPR2genes showed highest expression (P <0.01) in catagen primary follicles. It isnot obvious that the difference between the expression of anagen and telogen.(4) In addition, RT-PCR analysis showed that GMPR2gene was only present in goatskin, not in liver and heart.(5)GMPR2expression quantity decrease after liaoning new breeding cashmere goatsanagen cells melatonin treatment. Besids, the melatonin concentration of0.2g/L, treatmentfor48h, GMPR2gene expression levels decreased most significantly (P <0.01). (6)We analyses the antibody titer and specific identification by the ELISA method andWesten blot methods. The results show that the titer reached1:3.2million or more, andspecificity preferable.It meant that prepared anti-K26polyclonal antibody is successful.