南方医科大学学报

南方医科大学学报 ›› 2004, Vol. 24 ›› Issue (09): 1033-1036.

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精子基因表达谱研究初探

毛向明1, 马文丽1, 冯春琼1, 邹亚光2, 郑文岭3   

  1. 1. 第一军医大学, 分子生物学研究所, 广东, 广州, 510515;
    2. 第一军医大学, 南方医院口腔科, 广东, 广州, 510515;
    3. 广州军区广州总医院分子肿瘤研究所, 广东, 广州, 510010
  • 出版日期:2004-09-20 发布日期:2004-09-20
  • 基金资助:
    收稿日期:2004-3-26。
    基金项目:国家自然科学基金(39880032);广州市重点科技攻关项目(99-Z-022-01)
    作者简介:毛向明(1963-),男,第一军医大学在读博士研究生,讲师、主治医师,电话:020-85140114-89097,E-mail:mxm@fimmu.com

An initial examination of the spermatozoal gene expression profile

MAO Xiang-ming1, MA Wen-li1, FENG Chun-qiong1, ZOU Ya-guang2, ZHENG Wen-ling3   

  1. 1. 第一军医大学, 分子生物学研究所, 广东, 广州, 510515;
    2. 第一军医大学, 南方医院口腔科, 广东, 广州, 510515;
    3. 广州军区广州总医院分子肿瘤研究所, 广东, 广州, 510010
  • Online:2004-09-20 Published:2004-09-20

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摘要： 目的 应用限制性显示技术,收集正常成年男性的精子基因表达谱探针,制备相应的精子表达谱芯片,以对成年男性正常精子的基因表达情况有一全面、系统的了解。方法 提取成年男性正常精子的总RNA,逆转录为cDNA,经限制性内切酶酶切、加接头、PCR扩增、转化、克隆、鉴定等操作,收集成年男性正常精子的基因表达谱探针,制备相应的精子表达谱芯片,并进行初步的杂交分析。结果 收集了1 859个基因片段,取其中368个基因探针制备芯片,进行了小量初试。结论 证实在精子中有大量基因表达;提示我们自行制备的精子基因表达谱探针也具有较好的特异性和可信性。

Abstract: Objective To collect the normal spermatozoal gene expression sequence tags with the restriction display technique for constructing a microarray to understand spermatozoal gene expression profiles. Methods The total RNA extracted from normal human spermatozoa were reversely transcribed into cDNAs, which were digested by Sau3AI and linked to universal adapters (adapter 1) at both ends. According to the sequence of the adapter, a pair of primers (universal primers 1) was designed, followed by PCR with primers 1 and the PCR products were transferred into E.coli. The positive clones were collected after identification to serve as the probes for constructing the gene expression microarray of spermatozoa by printing those probes on the slides. The accomplished microarrays were examined by Cy3-labeled normal spermatozoal samples. Results Altogether 1 859 probes were collected, from which 368 were picked out randomly for constructing the microarray. Conclusions Human spermatozoa contain a rich repertoire of RNAs, and the probes we prepared possess good incredibility and speciality.

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