中国南瓜CmNPR1基因的克隆和表达分析

Cloning and Expression Analysis of CmNPR1 Gene in Cucurbita moschata

从实验室前期对中国南瓜雌花败育转录组测序结果推测,CmNPR1基因可能在南瓜花发育过程中发挥重要功能。该研究以中国南瓜自交系‘3-1’为试验材料,采用同源克隆方法获得中国南瓜CmNPR1基因CDS序列,通过生物信息学、基因表达以及亚细胞定位分析对该基因进行初步研究,为进一步研究CmNPR1基因在南瓜花发育中的功能和作用机制奠定基础。结果表明:(1)中国南瓜CmNPR1基因CDS全长1442 bp,编码480个氨基酸;蛋白序列包含有一个BTB/POZ和一个锚蛋白重复序列(Ank)保守结构域;该蛋白无信号肽和跨膜结构;多序列比对分析结果显示,CmNPR1氨基酸序列与美洲南瓜的亲缘关系最近,为96.05%,其次是印度南瓜,为95.63%。(2)CmNPR1基因在所取样品中花纵径0.5 cm时期表达量最高,且在花不同结构中柱头的表达量最高。(3)通过拟南芥原生质体亚细胞定位分析发现,该蛋白定位于细胞质和细胞核。

Based on the transcriptome sequencing of aborted female flower in Cucurbita moschata,it is suggested that the CmNPR1 may play an important role in flower development of C.moschata.In this study,we used homology cloning method to obtain the CDS of CmNPR1 from the inbred line‘3-1’.In order to provide a research foundation for further study on the function and mechanism of CmNPR1 in flower development of C.moschata,we analyzed the bioinformatics,expression characteristics and subcellular localization of CmNPR1.The results showed that:(1)the length of the coding sequence of CmNPR1 is 1442 bp,and it encodes a 480 amino acid-long protein.The protein sequence contains a BTB/POZ and an ankyrin repeats(ANK),and it has no signal peptide and transmembrane structure.The results of multiple sequence alignment showed that the CmNPR1 have the closest genetic relationship with Cucurbita pepo(96.05%),followed by Cucurbita maxima(95.63%).(2)The expression level of CmNPR1 was the highest at the minimum flower developmental stage(0.5 cm),and the expression level in stigma was the highest among different flower structures.(3)The subcellular localization analysis showed that the CmNPR1 was localized to the cytoplasm and nucleus.