Cloning And Expression Of Cashmere Goat PH-30 Gene

The mechanism of sperm-egg fusion is not only a significant theory of animal impregnation, but also have important prasense practical application of immunocontraception and immunity infertility. PH-30 gene is the best characterized molecule involved in the sperm-egg plasma membrane interaction ,as one of a few sperm membrane surface proteins which play vital role in binding and fusion of sperm-egg plasmalemma ,fertilinβis an important protein in many mammalian sperm membrane.In this study, we cloned goats PH-30 gene . First, extracted from goat testis total RNA, used as a template, synthesized cDNA through reverse transcription, designed a pair of primers according to four kinds of animals PH-30 cDNA sequence conservation area published by GenBank, and got DNA fragments of 448 bp by PCR amplification. Then, using RT-PCR and Northern hybridization detected the expression of PH-30at the mRNA level in different parts of the goat testis and epididymis.Finally we got goats PH-30 gene full-length cDNA sequences using RACE technology. The major results are as follows:1,The partial sequences of cashmere goat PH-30 cloned by PT-PCR. The nucleotide was sequenced and analyzed. 99% identity with sheep PH-30 and 92% homology with bovine PH-30 were found.2,Based on the known cDNA sequence of cashmere goat, cDNA 5 'and 3' end sequence of PH-30 we successfully cloned using RACE technology. the full-length.The full-length cDNA is 2611. One open reading frame (ORF) consisting of 2241 bp encoded 747 aa.3,Expression analysis by RT-PCR and Northern hybridization revealed that PH-30 was only expressed in the testis when compared to caput,corpus,cauda epididymis.