CORE--MICROINJECTION

将蛋白质和抗体显微注射到活的体细胞中， 一个强大的方法来研究调控组件的功能， 细胞。 这种方法在研究 癌基因蛋白及其在哺乳动物细胞生长控制中的功能， 在这种情况下，很难通过 任何其他手段。 通过引入功能性癌基因蛋白质， 可以以特定的方式刺激生长;而通过注射 抑制性抗体或寡核苷酸，细胞生长可以被特异性地 被挡出. 我们将继续利用高效的显微注射 在过去的项目期间建立的设施，以研究 通过蛋白质磷酸化调节核癌基因活性， 去磷酸化 除了微量注射特定的 中和抗体和反义寡核苷酸 生长因子信号转导系统的组成部分，我们将 显微注射各种信号蛋白和核癌蛋白。 这些 将包括cJun、cJun突变体、PP 2A催化和调节亚基， SH 2结构域，酪氨酸磷酸酶，E2 A-Pbx 1，vAb 1，Ha-Ras，酪蛋白激酶 II，erkI和erkII。 这些大分子对细胞的影响 将检测增殖和基因表达。

Microinjection of proteins and antibodies into living somatic cells is a powerful approach to studying the function of regulatory components of the cells. This method has been particularly useful in studies of oncogene proteins and their function in mammalian cell growth control, where it is difficult to alter the complement of expressed proteins by any other means. By introducing a functional oncogene protein, cell growth can be stimulated in a specific way; whereas by injection of an inhibitory antibody or oligonucleotide, cell growth can be specifically blocked. We will continue to utilize an efficient microinjection facility established during the past project period to study the regulation of nuclear oncogene activity by protein phosphorylation and dephosphorylation. In addition to microinjection of specific neutralizing antibodies and antisense oligonucleotides directed against components of the growth factor signal transduction system, we will microinject various signalling proteins and nuclear oncoproteins. These will include cJun, cJun mutants, PP2A catalytic and regulatory subunits, SH2 domains, tyrosine phosphatases, E2A-Pbx1, vAb1, Ha-Ras, casein kinase II, erkI and erkII. The effects of these macromolecules on cell proliferation and gene expression will be examined.