STRUCTURE AND EXPRESSION OF MAMMALIAN TUBULIN GENES

哺乳动物微管蛋白基因的结构和表达

• 批准号: 3280011
• 负责人: NICHOLAS COWAN
• 金额: $ 4.49万
• 依托单位: NEW YORK UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-09-01 至 1991-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3280011
• 关键词: DNA replication; antiserum; biological polymorphism; cell differentiation; cell free system; chickens; chromosome complement; chromosomes; cytoskeleton; deoxyribonuclease I; developmental genetics; gene expression; genetic library; genetic manipulation; genetic mapping; genetic transcription; haptens; human population genetics; immunogenetics; in situ hybridization; laboratory mouse; messenger RNA; microtubules; molecular cloning; neoplastic transformation; nucleic acid hybridization; nucleic acid probes; nucleic acid sequence; nucleoproteins; tissue /cell culture; tubulin

This proposal concerns the structure, evolution and expression of mammalian tubulin genes. These genes encode the Alpha- and Beta-subunits of microtubules, structures that perform an essential role in many vital organelles--cilia, flagella, the mitotic spindle and the cytoskeleton--of all eucaryotic cells. The proposed approach will involve 1) characterization by DNA sequence analysis of human Beta-tubulin genes that encode distinct Beta-tubulin isotypes. 2)a) Synthesis of peptide haptens corresponding to small carboxyterminal regions that are unique to individual Beta-tubulin isotypes, b) use of these haptens in hapten/carrier conjugates for the generation of Beta-tubulin isotype-specific monoclonal antisera, c) application of these sera in immunofluorescence experiments to determine whether two or more Beta-tubulin isotypes can coassemble in a single microtubule. 3)a) Isolation of cDNA clones encoding mouse Alpha- and Beta-tubulin sequences from cDNA libraries constructed from whole embryo and adult polyA+ mRNA, b) generation of gene-specific 3'-untranslated region subcloned probes, c) use of these probes in in situ hybridization experiments using cryostat sections of staged animals to study the tissue-specific developmental regulation of individual tubulin genes. 4) Use of Beta-tubulin gene/BPV shuttle vectors for the identification of sequences within human Beta-tubulin genes that are critical in determining changes in the level of cytoplasmic Beta-tubulin mRNAs that occur in response to modulation of the size of intracellular monomer pools. 5)a) Construction of cloned probes encoding Tau and MAP sequences, b) use of these probes i) to determine the number and evolutionary conservation of the sequences encoding these proteins, ii) to assess the response of Tau and/or MAP mRNA levels to changes in the level of tubulin monomer, iii) to study the developmental regulation of these proteins by in situ hybridization.

这一建议涉及哺乳动物的结构、进化和表达

项目成果

Diverse mechanisms in the generation of human beta-tubulin pseudogenes.

人类β-微管蛋白假基因产生的多种机制。

• DOI: 10.1126/science.6178164
• 发表时间: 1982
• 期刊: Science (New York, N.Y.)
• 作者: Wilde,CD;Crowther,CE;Cowan,NJ
• 通讯作者: Cowan,NJ

Structural variation among human beta-tubulin genes.

人类β-微管蛋白基因的结构变异。

• DOI: 10.1073/pnas.78.8.4877
• 发表时间: 1981
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: Cowan,NJ;Wilde,CD;Chow,LT;Wefald,FC
• 通讯作者: Wefald,FC

Structure of two human alpha-tubulin genes.

两个人类α-微管蛋白基因的结构。

• DOI: 10.1073/pnas.79.1.96
• 发表时间: 1982
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: Wilde,CD;Chow,LT;Wefald,FC;Cowan,NJ
• 通讯作者: Cowan,NJ

Isolation of a multigene family containing human alpha-tubulin sequences.

含有人α-微管蛋白序列的多基因家族的分离。

• DOI: 10.1016/0022-2836(82)90486-7
• 发表时间: 1982
• 期刊: Journal of molecular biology
• 影响因子: 5.6
• 作者: Wilde,CD;Crowther,CE;Cowan,NJ
• 通讯作者: Cowan,NJ