| 摘要: |
| 亲环素是一个多基因家族,在植物生命活动中发挥着重要的作用。该研究以大花杓兰(Cypripedium macranthum)为材料,采用RT-PCR技术克隆到1个亲环素基因(CyP),并对其进行生物信息学分析。结果表明:大花杓兰CyP基因的开放阅读框序列为525 bp,命名为CmCyP(GenBank登录号为MH411125),编码174个氨基酸。预测CmCyP蛋白是一个位于细胞质、相对分子量约为18 kD、理论pI为8.73、无信号肽、跨膜结构域的亲水性蛋白质。磷酸化和糖基化位点预测分析发现,CmCyP蛋白存在18个潜在的磷酸化位点和2个潜在的糖基化位点。蛋白保守结构域预测分析发现,CmCyP蛋白包含一个高度保守的肽脯氨酰顺反异构酶结构域,属于单结构域亲环素。对二级结构进行预测分析发现,CmCyP蛋白中存在无规卷曲70个、延伸链56个、α-螺旋23个、β-折叠25个,这4种结构元件在三级结构中也有体现。系统进化树结果显示,大花杓兰CmCyP蛋白与铁皮石斛(Dendrobium catenatum)和万带兰(Vanda hybrid cultivar)的CyP蛋白的亲缘关系较近。该研究首次克隆了大花杓兰亲环素基因(CmCyP),为进一步探讨CmCyP基因的生物学功能奠定了基础。 |
| 关键词: 大花杓兰, RT-PCR, 基因克隆, 系统发育树, 序列分析 |
| DOI：10.11931/guihaia.gxzw201803035 |
| 分类号:Q949.95 |
| 文章编号:1000-3142(2019)05-0633-08 |
| Fund project:国家自然科学基金(31100314); 河北省教育厅青年拔尖人才项目(BJ2016045); 廊坊师范学院博士基金(LSLB201405)[Supported by the National Natural Science Foundation of China(31100314); Talented Youth Program of Hebei Education Department(BJ2016045); Doctoral Foundation of Langfang Normal University(LSLB201405)]。 |
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| Cloning and bioinformatics analysis of cyclophilin gene (CmCyP)from Cypripedium macranthum |
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FU Yajuan1,2, ZHANG Jian3, LIU Huan1, HOU Xiaoqiang1,2*
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1. College of Life Sciences, Langfang Normal University, Langfang 065000, Hebei, China;2. Edible and Medicinal
Fungi Research and Development Center of Hebei Universities, Langfang 065000, Hebei, China;3. Langfang Polytechnic Institute, Langfang 065000, Hebei, China
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| Abstract: |
| Cyclophilin is a multi-gene family that plays an important role in plant biology. A cyclophilin gene in Cypripedium macranthum named as CmCyP (GenBank Accession No. MH411125), was successfully cloned by RT-PCR, and then bioinformatics analysis were carried out. The results were as follows: CmCyP protein was predicted as a stable hydrophilic protein composed of 174 amino acids, without signal peptide and transmembrane domain. Its relative molecular mass was approximately 18 kD, and a predicated isoelectric point(pI)was 8.73.Phosphorylation and glycosylation site prediction analysis revealed that eighteen potential phosphorylation sites and two potential glycosylation sites were found in CmCyP protein. Protein domain prediction revealed that CmCyP contained a highly conserved peptide prolyl cis-trans isomerase domain, belonging to single domain of cyclophilin. The secondary structure of CmCyP was abundant in random coils(70)and extension chains(56), while was less in α-helices(23)and β-turns(25). Phylogenetic tree analysis indicates that the genetic relationship of CmCyP in C. macranthum with CyPs from Dendrobium catenatum and Vanda hybrid cultivar is relatively close. In conclusion, the CmCyP gene was firstly cloned from Cypripedium macranthum, which provides for further study on biology function of CmCyP gene. |
| Key words: Cypripedium macranthum, RT-PCR, gene cloning, phylogenetic tree, sequence analysis |
