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Identification of differentially expressed genes of primary spermatocyte against round spermatid isolated from human testis using the laser capture microdissection technique

摘要The method of laser capture microdissection (LCM) combined with suppressive subtractive hybridization (SSH) was developed to isolate specific germ cells from human testis sections and to identify the genes expressed during differentiation and development. In the present study, over 10,000 primary spermatocytes and round spermatid cells were successfully isolated by LCM. Using the cDNAs from primary spermatocytes and round spermatids, SSH cDNAs library of primary spermatocyte-specific was constructed. The average insert size of the cDNA isolated from 75randomly picked white clones was 500 bp, ranging from 250 bp to 1.7 kb. Using the dot-blot method, a total of 421clones were examined, resulting in the identification of 390 positive clones emitting strong signals. Partial sequence of cDNAs prepared from each clone was determined with an overall success rate of 84.4%. Genes encoding cytochrome c oxidase Ⅱ and the rescue factor-humanin were most frequently expressed in primary spermatocytes, suggesting their roles involved in meiosis.

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作者单位 National Laboratory Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences,Peking Union Medical College, 5 Dong Dan San Tiao, 100005 Beijing, China;Chinese National Human Genome Center, Beijing, 3-707 North Yon [1] National Laboratory Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences,Peking Union Medical College, 5 Dong Dan San Tiao, 100005 Beijing, China [2] National Research Institute for Family Planning, WHO Collaboration Center for Research in Human Reproduction, Beijing, 12 Da HuiSi, 100081 Beijing, China [3] Center for Biomedical Research, Population Council, 1230 York Avenue, New York, NY 10021, USA [4]
分类号 Q2
栏目名称 ARTICLES
DOI 10.1038/sj.cr.7290254
发布时间 2005-05-26
基金项目
国家自然科学基金 国家高技术研究发展计划(863计划) 国家重点基础研究发展计划(973计划)
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细胞研究(英文版)

细胞研究(英文版)

2004年14卷6期

507-512页

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