| 张恒宇,柳逸群,葛源.莱茵衣藻type I metacaspase基因克隆及其参与调控程序性细胞死亡研究.渔业科学进展,2019,40(6):131-137 |
| 莱茵衣藻type I metacaspase基因克隆及其参与调控程序性细胞死亡研究 |
| Cloning of the Metacase Type I Gene of Chlamydomonas reinhardtii and Its Involvement in the Regulation of Programmed Cell Death |
| 投稿时间：2018-05-07 修订日期：2018-05-17 |
| DOI：10.19663/j.issn2095-9869.20180507002 |
| 中文关键词: Metacaspase 莱茵衣藻 程序性细胞死亡 |
| 英文关键词: Metacaspase Chlamydomonas reinhardtii Programmed cell death |
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| 中文摘要: |
| Metacaspase是在原生动物、真菌和植物中发现的一种具有底物精氨酸/赖氨酸特异性的半胱氨酸蛋白酶。根据蛋白质结构特征可将metcaspase分为typeⅠ与typeⅡ两种类型，均参与调控多种植物与原生动物程序性细胞死亡。本研究根据GenBank数据库莱茵衣藻(Chlamydomonas reinhardtii) type I metacaspase基因(GenBank No. XM_001696904)序列，采用巢式PCR克隆获取type I metcaspase基因开放阅读框(ORF)序列并命名为CrMC1。CrMC1 ORF全长987 bp，推测编码1个包含405个氨基酸的蛋白质。通过与已知物种type I metacaspase氨基酸序列进行同源序列比对发现，CrMC1具有高度保守的p20、p10、连接区结构域以及精氨酸、半胱氨酸活性中心位点。研究显示，在H2O2诱导的莱茵衣藻程序性细胞死亡过程中，CrMC1表达量显著提高(P<0.05)，2 h后达到峰值，3 h时下降至对照组同一水平。结果表明，莱茵衣藻type I metacaspase基因CrMC1参与H2O2诱导的莱茵衣藻程序性细胞死亡调控。 |
| 英文摘要: |
| The metacaspase is a cysteine protease with the substrate of arginine/lysine, specifically found in protozoa, fungi, and plants. The metacaspase is generally believed to be closely related to the caspase of the metazoan. Previous studies have shown that metacaspases can be divided into type I and typeⅡ according to the differences in their structural characteristics, and both types have been found to be involved in the regulation and the control of programmed cell death in various plants and protozoa. Based on the information of the sequence of the Chlamydomonas reinhardtii type I metacaspase cDNA, obtained from the Genbank database (Genbank NO. XM_001696904), this study was able to use the method of two-step nested PCR cloning to retrive the Open Reading Frame (ORF) of the sequence of the C. reinhardtii type I metacaspase cDNA. The ORF sequence, mentioned above, was named as the CrMC1 in this study. The length of the CrMC1 ORF, in its entirety, is 987 bp, from which can be inferred that there are 405 amino acids encoded in this sequence. By the method of homologous sequence comparative analysis with the already known type I metacaspase protein, it was then found that the CrMC1 has conservative p20, p10, linker domain, and arginine and cysteine active center sites. Furthermore, this study is also able to show that, during the H2O2 induced programmed cell death of C. reinhardtii, the gene transcripts of the CrMC1 was increased at a statistically significant level (P<0.05). The gene transcripts of the CrMC1 then reached its peak after 2 h. Eventually, the gene transcripts of the CrMC1 decreased to the same level as it was in the control group at 3 h. In conclusion, the results as discussed above have indicated the following: the C. reinhardtii CrMC1 was involved in the regulation and the control of the H2O2 induced programmed cell death of C. reinhardtii. |
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