mRNA差异显示法克隆原发无精症相关基因

目的克隆与原发无精症发病相关的基因。方法运用改进的ｍＲＮＡ差异显示技术，从原发无精症患者和正常成人睾丸组织中获得两者的差异表达片段，即表达序列标志（ｅｘｐｒｅｓｓｅｄｓｅｑｕｅｎｃｅ－ｔａｇ，ＥＳＴ），并克隆测序分析。结果正常成人和原发无精症患者的睾丸组织存在明显的基因表达差异。所分析的５个在正常成人睾丸中表达、而在患者组织中不表达的差异片段中，１个与ＧｅｎＢａｎｋ中ｃｏｓｍｉｄＬ２７ｈ９的部分序列１００％同源，该ｃｏｓｍｉｄ插入序列位于４ｐ１６．３的亨廷顿舞蹈病基因区（Ｈｕｎｔｉｎｇｔｏｎ＇ｓｄｉｓｅａｓｅｒｅｇｉｏｎ），另４个为无同源序列的新基因片段。结论４ｐ１６．３的亨廷顿舞蹈病基因区可能与原发无精症相关，并在睾丸中表达。

Cloning of the idiopathic azoospermia related genes with mRNA differential display method

bjective Defect of some genes related to spermatogenesis may result in male infertility with azoospermia. This study aimed to clone some of the genes. Methods Testis tissues from a patient with idiopathic azoospermia and from a normal fathered man were investigated with an improved mRNA differential display approach. Results Significant difference was observed between the two tissues in gene expression.Five differentially expressed sequencetags(ESTs) were cloned and sequenced. Homology analysis with software advanced BLAST 2.0 showed that one EST shared 100% homology with cosmid L27h9, which located in Huntington's disease region on 4p16.3; the other 4 showed very low homology with sequences in GenBank. Conclusion Azoospermia has complex genetic heterogeneity. A gene located in Huntington's disease region on 4p163 is expressed in human testis during the spermatogenesis, and a loss of its function may associate with azoospermia.