关键词: HSL 基因, SRY 基因, 精子, 荧光原位PCR, FITC( 异硫氰酸荧光素)

Abstract: Abstract: Purpose Spermatogenesis is a fine control process with mitosis and meiosis， which is from spermatogonial stem cells began in male seminiferous tubule． By the phenomenon proportion of made to female in mammals，which is 1 ∶ 1，we can infer that the fertility rate of sperm with X gene and Y gene should be 1 ∶ 1． ·4· 第1 期朱璇等: 在单细胞水平建立原位PCR 基因鉴别技术平台———以精子基因型鉴别为例 However，after differentiation，X and Y sperm for 1 ∶ 1 ratio is not known through spermatogenesis． Not all spermatocytes can develop into sperm during spermatogenesis eventually． However，to study the male and female birth ratio need to first study that if the spermatogenesis is strictly in accordance with 1∶ 1． Therefore，the present study，using a rapid and reliable platform of In-situ PCR， can further support the studies on regulation in spermatogenesis． Step Using HSL gene exploration steps，we can obtain an optimal fluorescence in-situ PCR experiment scheme，including proteinase K effect time and concentration，sperm depolymerization method and degree，etc，then applies to SRY gene． Result By Nikon fluorescence microscopy，we can clearly see the sperm head with fluorescent signals． Sperm head with green fluorescent signals target to SRY gene is the Y chromosome sperm ( Y sperm) ． Our lab tested 493 Kunming mice sperm altogether and there were total 273 signals． X sperm ∶ Y sperm = 55. 37% ． The experimental results approached to draw after the chi-square test． Conclusion We can use in-situ PCR technology to quickly appraisal the sperm“gender”in single-celled level，which is also suitable to detect any kind of gene in whatever the cell type is．

Key words: HSL gene, SRY gene, sperm, fluorescence in-situ PCR;