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小鼠卵母细胞中独特的蛋白激酶C谱:逆转录聚合酶链反应和蛋白质免疫印迹显示缺乏钙依赖性传统亚型

Unique protein kinase C profile in mouse oocytes: lack of calcium-dependent conventional isoforms suggested by rtPCR and Western blotting.

作者信息

Gangeswaran R, Jones K T

机构信息

Medical Research Council Experimental Embryology and Teratology Unit, St. George's Hospital Medical School, London, UK.

出版信息

FEBS Lett. 1997 Jul 28;412(2):309-12. doi: 10.1016/s0014-5793(97)00782-5.

Abstract

rtPCR and Western blotting were used to determine which members of the PKC family are present in both immature and mature mouse oocytes. Using isoform-specific PCR primers and antibodies PKC-delta and -lambda were detected while such techniques failed to observe the conventional isoforms of PKC-alpha, -beta, -gamma. This isoform profile was confirmed using an alternative PCR strategy, which allowed discrimination of PCR products derived from conventional and novel PKC isoforms. In addition PKC-epsilon, -eta, -theta and -zeta were not detected by rtPCR. These results suggest that the predominant isoforms in oocytes are PKC-delta and -lambda.

摘要

逆转录聚合酶链反应(rtPCR)和蛋白质免疫印迹法被用于确定蛋白激酶C(PKC)家族的哪些成员存在于未成熟和成熟的小鼠卵母细胞中。使用亚型特异性的PCR引物和抗体检测到了PKC-δ和-λ,而这些技术未能观察到PKC-α、-β、-γ的传统亚型。使用另一种PCR策略证实了这种亚型分布,该策略能够区分源自传统和新型PKC亚型的PCR产物。此外,rtPCR未检测到PKC-ε、-η、-θ和-ζ。这些结果表明,卵母细胞中的主要亚型是PKC-δ和-λ。

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