HELICASE GENE PL10 IN MOUSE GERM CELL DEVELOPMENT

解旋酶基因 PL10 在小鼠生殖细胞发育中的作用

基本信息

批准号：
2471394
负责人：
DONNA R SESSION
金额：
$ 8.14万
依托单位：
MAYO CLINIC ROCHESTER
依托单位国家：
美国
项目类别：
财政年份：
1992
资助国家：
美国
起止时间：
1992-07-01 至 1998-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/2471394
关键词：
chimeric proteins gene expression genetic library genetic promoter element genetic transcription genetic translation genetically modified animals germ cells helicase introns laboratory mouse molecular cloning northern blottings nucleic acid sequence nucleic acid structure restriction mapping spermatogenesis yeasts

项目摘要

The murine gene PL10 was isolated in a search for genes that may play a role in the regulation of spermatogenesis. It is a member of the newly defined helicase gene family. One function proposed for members of the helicase gene family is the regulation of gene expression through translational control. Phase I of this proposal will include research to further characterize the PL10 gene, its expression and protein products, as well as specific courses. Phase II will be dedicated to identifying the function of PL10. Initially, a testis cDNA library will be screened for full length cDNAs corresponding to the 3.2 and 3.7 kb testis-specific PL10 transcripts. The differences between these two transcripts will be explored through restriction mapping and sequencing. The full length cDNAs will then be used to synthesize the PL10 protein(s) in vitro and generate antibodies against PL10. The antibodies will be used to determine temporal, tissue, cellular specificity and intracellular location of the PL10 protein product. If the 3.2 and 3.7 kb transcripts code for different proteins, specific antibodies will be generated to these proteins to further explore the differences in the two gene products. A mouse genomic library will be screened for the PL10 gene. PL10's gene organization will be characterized via restriction mapping, S1 analysis and sequencing intron-exon boundaries. In Phase II, two different strategies will be employed to identify the function of the PL10 gene. One method will be to experimentally manipulate the expression of the PL10 gene in transgenic animals. The second method will involve complementation analysis of the yeast homolog of PL10. This research experience will provide the necessary knowledge and skills to become an independent researcher and apply this experience to the fields of developmental genetics and reproductive endocrinology.

在搜索可能发挥的基因时，将鼠基因PL10分离出来在精子发生调节中的作用。它是新的成员定义的解旋酶基因家族。为成员提出的一个功能解旋酶基因家族是通过基因表达的调节翻译控制。该提案的第一阶段将包括研究进一步将PL10基因（其表达和蛋白质产物）作为以及特定的课程。第二阶段将致力于确定 PL10的功能。最初，将筛选睾丸cDNA库对应于3.2和3.7 kb睾丸特异性PL10的全长cDNA 成绩单。这两个成绩单之间的差异将是通过限制映射和测序探索。全长cdnas 然后将用于在体外合成PL10蛋白并生成针对PL10的抗体。抗体将用于确定时间，组织，细胞特异性和细胞内位置 PL10蛋白产物。如果3.2和3.7 kb成绩单代码不同蛋白质，特异性抗体将与这些蛋白质生成进一步探索两个基因产物的差异。小鼠基因组库将筛选为PL10基因。 PL10的基因组织将通过限制映射，S1分析和测序来表征内含子 - 外观边界。在第二阶段，将有两种不同的策略用于识别PL10基因的功能。一种方法是实验操纵PL10基因在转基因中的表达动物。第二种方法将涉及互补分析 PL10的酵母同源物。这项研究经验将提供必要的知识和技能成为一名独立研究人员并应用此发展遗传学和生殖领域的经验内分泌学。