Constracation And Study On Male-special Genes CDNA Subtractive Library Of Mouse Cerebrum

Male-specific antigen is usually defined as a male minor histocmpatibility Y antigen, which was confirmed by Eichwald and Silmser in 1955. Male-specific antigen is of great meaning in studing on sex differentiation, sex control and transplantation between male and female animals.A subtractive cDNA Library of Cerebrum of BALA/C mouse was constracted using suppression subtractive hybridization. Methods: Total RNA was isolated from Cerebrum of Male and female BALA/C mouse. mRNA was extracted and converted into double-strand cDNA. Male-specific cDNA was obtained by Using suppression subtractive hybridization ( male mouse as driver, female mouse as tester) and cloned into pGEM-T Vector, then the vector was transformed into E. coli JM109. Finally, 160 clones were obtained from subtracted cDNA Libraries. By PCR detection, the number of positive clones was 134 and the length of the inserted fragments of subtracted cDNA library was between 0.2 and 0.8kb. The positive rate is 83.7%. 80 clones were analyzed by Blastn analysis in Genebank nr database. Result showed that 3 sequences was novel sequence and 77 sequences have homology with EST in database. 3 novel sequences were blast analyzed in MGI batabase. Result showed that these sequences locus in 6 chromosome.The results showed that the subtracted libraries constructed were suitable for further study on the male-specific antigen of mouse, which provid a novel study method and a novel technical system. It will beneficial to studing on sex difference, sex control and sexual related graft rejection.