QUANTITATION OF AMYLASE MRNA BY CDNA HYBRIDIZATION

通过 CDNA 杂交对淀粉酶 mRNA 进行定量

• 批准号: 3425218
• 负责人: SUN-KEE KIM
• 金额: $ 2.07万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-05-01 至 1989-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3425218
• 关键词: aging; amylases; complementary DNA; exocrine glands; genetic transcription; genetic translation; glucocorticoids; messenger RNA; parotid gland

This research program is concerned with regulation of secretory protein synthesis in exocrine glands, and changes in the regulatory mechanism during aging. The immediate goal is to determine whether secretory protein synthesis is regulated at the level of transcription or translation. In rat parotid glands, changes in rates of secretory protein synthesis in relation to aging, secretory (discharge) activity and varying levels of glucocorticoids accompany parallel changes in translational activities of total messenger RNA (mRNA) and mRNA specific for amylase, a major secretory protein of the gland. It is unknown if the changes in messenger activities are due to actual changes in the amount of mRNA involving regulation at the level of transcription. Quantitations of mRNA specific for secretory protein are possible using a cloned amylase complementary DNA (cDNA) probe, since amylase makes up a large proportion of the secretory proteins in parotid glands. cDNA clones, pucg, containing copies of salivary amylase sequence at pst 1 sites have been provided for this study by Dr. Miriam H. Meisler of this University. Specific aims are: 1. To establish technical procedures for amylase cDNA probe generation and hybridization for quantitation of amylase mRNA by dot blot and northern blot analyses. 2. To test the feasibility of applying the above techniques to determine if the increased rate of amylase synthesis following stimulated secretion of stored secretory proteins is accompanied by a parallel increase in the amount of amylase mRNA.

这个研究项目是关于分泌蛋白的调节