干旱区生态环境知识资源中心(Arid): 干旱胁迫条件下发菜cphB差异表达与基因克隆

Arid

	干旱胁迫条件下发菜cphB差异表达与基因克隆
其他题名	Differential Expression and Gene Cloning of cphB from Nostoc flagelliforme under Drought Stress
	王淑萍; 刘阳; 严奉坤; 吴诗杰; 王玲霞; 徐婷婷; 梁文裕
来源期刊	基因组学与应用生物学
ISSN	1674-568X
出版年	2017
卷号	36 期号:6 页码:2472-2479
中文摘要	发菜是一种分布在干早、半干旱荒漠草原地区的陆生固氮蓝藻,对干旱具有极强的适应性。采用qRT-PCR技术,对干旱胁迫条件下发菜cphB的差异表达进行了分析,发现干旱胁迫条件下发菜cphB在转录水平上呈逐渐增加的趋势。根据发菜同源物种设计简并性引物克隆cphB基因,获得长度为864 bp的DNA (GenBank登陆号为KX092456)。同源性比较发现发菜cphB基因具有较高的保守性,二级结构预测表明发菜CphB由alpha螺旋,beta折叠和随机卷曲构成。三级结构预测表明CphB具有一个由alpha螺旋和beta折叠组成的5股的结构域和由7个beta折叠连成三明治结构的7股的结构域。将cphB基因在大肠杆菌中表达,获得符合预期的外源重组蛋白,CphB亚细胞定位于细胞质中。研究成果为进一步研究干旱胁迫条件下发菜cphB表达调控、藻蓝素的代谢机制及能量代谢机制方面奠定了基础。
英文摘要	Nostoc flagelliforme is a terrestrial nitrogen-fixing cyanobacte ria distributed in dry and semi arid desert steppe areas, which has strong adaptability to drought. We used qRT-PCR technique to analyze the differential expression of cphB from Nostoc flagelliforme under drought stress conditions and found that cphB gradually increased at the transcription level under drought stress. Degenerate primers were designed according to homologous species of Nostoc flagelliforme to clone cphB gene, and a DNA with the full length of 864 bp was obtained(GenBank access number: KX092456). Homology comparison showed that cphB gene had high conservatism, and the secondary structure prediction indicated that CphB was made up of alpha Helix, beta sheet and random coil. Besides, tertiary structure prediction demonstrated that CphB had a 5-stranded domain that composed of alpha Helix and beta sheet and a seven-stranded domain composed of 7 beta sheet with the structure like sandwich. The cphB was expressed in Escherichia coli, and the desired exogenous recombinant protein was obtained. In addition, CphB subcellular was located in the cytoplasm. The results laid a foundation for further study on expres sion and regulation of cphB, metabolic mechanism of phycocyanin and energy metabolism in Nostoc flagelliforme under drought stress.
中文关键词	发菜 ; 差异表达 ; 基因克隆 ; 原核表达
英文关键词	CphB Nostoc flagelliforme CphB Differential expression Gene clone Prokaryotic expression
语种	中文
国家	中国
收录类别	CSCD
WOS类目	PLANT SCIENCES
WOS研究方向	Plant Sciences
CSCD记录号	CSCD:6014472
来源机构	宁夏大学
资源类型	期刊论文
条目标识符	http://119.78.100.177/qdio/handle/2XILL650/236216
作者单位	宁夏大学生命科学学院, 银川, 宁夏 750021, 中国
推荐引用方式 GB/T 7714	王淑萍,刘阳,严奉坤,等. 干旱胁迫条件下发菜cphB差异表达与基因克隆[J]. 宁夏大学,2017,36(6):2472-2479.
APA	王淑萍.,刘阳.,严奉坤.,吴诗杰.,王玲霞.,...&梁文裕.(2017).干旱胁迫条件下发菜cphB差异表达与基因克隆.基因组学与应用生物学,36(6),2472-2479.
MLA	王淑萍,et al."干旱胁迫条件下发菜cphB差异表达与基因克隆".基因组学与应用生物学 36.6(2017):2472-2479.