| 摘要: |
| 通过对白及（Bletilla striata （Thunb.） Rchb.f.）的IRA1多拷贝抑制子（multicopy suppressor of IRA1，MSI1）基因序列进行生物信息学分析，并根据其核苷酸序列设计简单重复序列（simple sequence repeats， SSR）引物，进一步验证其在多个白及地方种中高度的保守性。运用Protparam、SOPMA、SWISS-MODEL等生物信息学软件分析白及MSI1蛋白的理化性质和结构域；DNAMAN、MEGA软件分别进行氨基酸多序列比对与系统进化分析；分析序列中的SSR位点并对多个白及品系进行PAGE检测和验证。结果显示，克隆得到的BSMSI1基因全长为3 700 bp，共编码了601个氨基酸残基，预测蛋白质分子量为65 309.75 kg/moL，等电点为5.95，表现出高度的保守性，而且发现不同地区的白及BSMSI1基因在遗传上具有多样性。本研究拓展了对 BSMSI1序列的认识，新开发标记能有效应用于MSI1基因的检测和育种鉴定，同时也可为其他物种的MSI1基因研究提供参考。 |
| 关键词: 白及 MSI1基因 结构与功能 生物信息学分析 简单重复序列 |
| DOI：10.14188/j.ajsh.2021.03.003 |
| 分类号:S567.239 |
| 基金项目:国家自然科学基金项目（31560079；31960074）；贵州省科学技术基金项目（黔科合平台人才［2017］5733-050，黔科合平台人才［2017］5712）；贵州省中医药管理局课题（QZYY-2019-060） |
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| Sequence analysis and SSR polymorphism analysis of MSI1 gene in Bletilla striata |
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HUANG Ceyin1, ZHAO Yongbin2, CHEN Ronghui1, LI Lin1, WEN Weie1, XU Delin1
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1.Department of Cell Biology， Zunyi Medical University， Zunyi 563099，Guizhou， China;2.Zhuhai Campus of Zunyi Medical University， Zhuhai，519041，Guangdong， China
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| Abstract: |
| By analyzing the sequences information of multicopy suppressor of IRA1 in Bletilla striata （BSMSI1）， simple sequence repeats （SSR） primers were designed based on the nucleotide sequence to further verify the high conservation in multiple Bletilla striata local species. The physical and chemical properties and domains of BSMSI1 protein were determined by a series of bioinformatics tools such as Protparam， SOPMA， SWISS-MODEL. Multiple alignment and phylogenetic tree were respectively achieved by DNAMAN. SSR sites in the sequence were analyzed and the PAGE electrophoresis confirmed that this primer pair had polymorphism. The sequence of BSMSI1 was 3 700 bp in length which encoded a protein of 601-amino-acid with a molecular weight of 65 309.75 kg/mol and an isoelectric point （pI） of 5.95. And this sequence showed a high degree of conservatism. Moreover， the genes of Bletilla striata in different regions were found to be genetically diverse. The results expanded the understanding of BSMSI1 gene sequence. The newly developed marker could be effectively applied to the detection of MSI1 gene and breeding projects， and it also provided reference for MSI1 researches of other species. |
| Key words: Bletilla striata MSI1 structure and function bioinfomatic analysis simple sequence repeats （SSR） |
