Chen Pan-Pan, Guo Qiao-Sheng, Shi Hong-Zhuan, Guo Li-Yuan, Zhang Hui-Ning
Institute of Chinese Medicinal Materials, Nanjing Agricultural University Nanjing 210095, China.
Zhongguo Zhong Yao Za Zhi. 2021 Feb;46(3):599-604. doi: 10.19540/j.cnki.cjcmm.20201114.102.
Protein kinase C(PKC) is a type of protein kinase widely involved in cell proliferation and development, but the developmental mechanism in the gonads of androgynous animals is still unclear. In order to explore the role of protein kinase C in the development of Whitmania pigra germ cells, the Wh. pigra PKC(Wp-PKC) gene was cloned, bioinformatics analysis was conducted, and fluorescent quantitative PCR was used to analyze the expression of female and male gonads. The results showed that:(1)The cloned Wp-PKC had a full length of 2 580 bp, a relative molecular weight of 76 555.19, and contains an open reading frame encoding 670 amino acids, Wp-PKC was closely related to Danio rerio PKC-α and rat PKC-γ. The similarity of amino acid sequence was 55% and 58%.(2)The protein encoded by Wp-PKC had no signal peptide and was a hydrophilic protein. The secondary structure is mainly composed of random coils, α-helices, extended chains, folds and folds, with the largest proportion of random coils and α-helices. Wp-PKC protein does not contain a transmembrane domain. Multiple sequence alignment and domain prediction analysis show that Wp-PKC contains 4 conserved domains of classical protein kinase C.(3)Fluorescence quantitative results showed that the expression of Wp-PKC in Wh. pigra gonads was positively correlated with the development of germ cells, and the expression in male gonads was significantly higher than that in female gonads. In summary, Wp-PKC is a classic PKC, and Wp-PKC may promote the development of Wh. pigra, especially the development of male gonads, and provide references for further research on the developmental mechanisms of Wh. pigra.
蛋白激酶C（PKC）是一种广泛参与细胞增殖和发育的蛋白激酶，但雌雄同体动物性腺中的发育机制仍不清楚。为了探究蛋白激酶C在宽体金线蛭生殖细胞发育中的作用，克隆了宽体金线蛭PKC（Wp-PKC）基因，进行了生物信息学分析，并采用荧光定量PCR分析了雌雄性腺中的表达情况。结果表明：（1）克隆得到的Wp-PKC全长2 580 bp，相对分子质量为76 555.19，包含一个编码670个氨基酸的开放阅读框，Wp-PKC与斑马鱼PKC-α和大鼠PKC-γ亲缘关系较近，氨基酸序列相似性分别为55%和58%。（2）Wp-PKC编码的蛋白无信号肽，是一种亲水性蛋白。二级结构主要由无规卷曲、α螺旋、延伸链、折叠和褶皱组成，其中无规卷曲和α螺旋所占比例最大。Wp-PKC蛋白不含跨膜结构域。多序列比对和结构域预测分析表明，Wp-PKC含有经典蛋白激酶C的4个保守结构域。（3）荧光定量结果显示，Wp-PKC在宽体金线蛭性腺中的表达与生殖细胞的发育呈正相关男性性腺中的表达明显高于女性性腺中的表达。综上所述，Wp-PKC是一种经典的PKC，Wp-PKC可能促进宽体金线蛭的发育，尤其是雄性性腺的发育，为进一步研究宽体金线蛭的发育机制提供参考。
