Goodnight J, Kazanietz M G, Blumberg P M, Mushinski J F, Mischak H
Molecular Genetics Section, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892.
Gene. 1992 Dec 15;122(2):305-11. doi: 10.1016/0378-1119(92)90219-f.
A 2199-bp complementary DNA (cDNA) that encodes protein kinase C-zeta (PKC-zeta) has been isolated from mouse brain by a combination of reverse transcription and primer extension. The predicted PKC-zeta protein consists of 592 amino acids which are 99% identical to those of rat PKC-zeta. Northern blots that were probed with this cDNA revealed abundant 2200-nucleotide (nt) and 4200-nt PKC-zeta mRNAs in mouse brain in roughly equal amounts. PKC-zeta mRNA was also abundant in normal lung, kidney, and testes, and in several hemopoietic tumor lines. In all other mouse tissues and cell lines that were examined, at least faint levels of PKC-zeta mRNAs could also be detected. In tissues other than brain, the amount of PKC-zeta mRNA was less, and the smaller species generally predominated. Furthermore, in these tissues, both PKC-zeta mRNAs appear to be approximately 200 nt longer than the two mRNAs found in the brain. When the cDNA is expressed in insect cells via a baculovirus expression vector, a 75-kDa protein is synthesized which, unlike other PKC isoforms, does not bind phorbol ester, even at very high concentrations.
通过逆转录和引物延伸相结合的方法，从小鼠脑中分离出了一段编码蛋白激酶C-ζ（PKC-ζ）的2199个碱基对的互补DNA（cDNA）。预测的PKC-ζ蛋白由592个氨基酸组成，与大鼠PKC-ζ的氨基酸序列有99%的同源性。用该cDNA进行Northern杂交分析发现，小鼠脑中存在大量2200个核苷酸（nt）和4200 nt的PKC-ζ mRNA，二者含量大致相等。PKC-ζ mRNA在正常肺、肾、睾丸以及几种造血肿瘤细胞系中也很丰富。在所有其他检测的小鼠组织和细胞系中，也至少能检测到微弱水平的PKC-ζ mRNA。在脑以外的组织中，PKC-ζ mRNA的含量较少，且较小的mRNA种类通常占主导。此外，在这些组织中，两种PKC-ζ mRNA似乎都比在脑中发现的两种mRNA长约200 nt。当通过杆状病毒表达载体在昆虫细胞中表达该cDNA时，会合成一种75 kDa的蛋白，与其他PKC同工型不同，即使在非常高的浓度下，该蛋白也不结合佛波酯。
