干旱区生态环境知识资源中心(Arid): Characterization of two copper/zinc superoxide dismutases (Cu/Zn-SODs) from the desert beetle Microdera punctipennis and their activities in protecting E. coli cells against cold

Arid

DOI	10.1016/j.cryobiol.2019.03.006
	Characterization of two copper/zinc superoxide dismutases (Cu/Zn-SODs) from the desert beetle Microdera punctipennis and their activities in protecting E. coli cells against cold
	Xikeranmu, Zilajiguli; Abdunasir, Maimaitiaili; Ma, Ji; Tusong, Kuerban; Liu, Xiaoning
通讯作者	Liu, Xiaoning
来源期刊	CRYOBIOLOGY
ISSN	0011-2240
EISSN	1090-2392
出版年	2019
卷号	87 页码:15-27
英文摘要	Superoxide dismutases (SODs) are crucial in scavenging reactive oxygen species (ROS); however, studies regarding SOD functions in insects under cold conditions are rare. In this paper, two novel Cu/Zn-SOD genes in the desert beetle Microdera punctipennis, an extracellular copper/zinc SOD (MpecCu/Zn-SOD) and an intracellular copper/zinc SOD (MpicCu/Zn-SOD), were identified and characterized. The results of quantitative real-time PCR showed that MpecCu/Zn-SOD expression was significantly up-regulated by 4 degrees C exposure for 0.5 h, but MpicCu/Zn-SOD was not. Superoxide anion radical (O-2(center dot-)) content in beetles under 4 degrees C exposure for 0.5 h showed an initial sharp increase and fluctuated during the cold treatment period, which was consistent with the relative mRNA level of MpecCu/Zn-SOD. The total SOD activity in the beetle was negatively correlated to the O-2(center dot-) content with a correlation coefficient of -0.437. An E. coli system was employed to study the function of each MpCu/Zn-SOD gene. The fusion proteins Trx-His-MpCu/Zn-SODs were over expressed in E. coli BL21 using pET32a vector, and identified by SDS-PAGE and Western blotting. The transformed bacteria BL21(pET32a-MpecCu/Zn-SOD) and BL21(pET32a-MpicCu/Zn-SOD) showed increased cold tolerance to -4 degrees C as well as increased SOD activity compared to the control BL21(pET32a). The relative conductivity and malondialdehyde content in the two MpCu/Zn-SODs transformed bacteria under -4 degrees C were significantly lower than the control BL21(pET32a). Furthermore, BL21(pET32a-MpecCu/Zn-SOD) had significantly higher SOD activity and cold tolerance than BL21(pET32a-MpicCu/Zn-SOD) under -4 degrees C treatment, and had lower conductivity than BL21(pET32a-MpicCu/Zn-SOD). In conclusion, low temperature led to the accumulation of O-2(center dot-) in M. punctipennis, which stimulated the expression of extracellular MpCu/Zn-SOD gene and the increase of total SOD activity. E. coli overexpressing Trx-His-MpCu/Zn-SODs increased resistance to cold treatment-induced oxidative stress. Our findings will be helpful in further study of Cu/Zn-SOD genes in insect cold-tolerance.
英文关键词	Microdera punctipennis Superoxide dismutase Cold stress Superoxide radicals E. coil Recombinant enzyme Antioxidant defense
类型	Review
语种	英语
国家	Peoples R China
收录类别	SCI-E
WOS记录号	WOS:000465154600002
WOS关键词	EXPRESSION ANALYSIS ; LIPID-PEROXIDATION ; GENE ; STRESS ; SYSTEMS ; CLONING
WOS类目	Biology ; Physiology
WOS研究方向	Life Sciences & Biomedicine - Other Topics ; Physiology
来源机构	新疆大学
资源类型	期刊论文
条目标识符	http://119.78.100.177/qdio/handle/2XILL650/215001
作者单位	Xinjiang Univ, Xinjiang Key Lab Biol Resources & Genet Engn, Coll Life Sci & Technol, 666 Shengli Rd, Urumqi 830046, Peoples R China
推荐引用方式 GB/T 7714	Xikeranmu, Zilajiguli,Abdunasir, Maimaitiaili,Ma, Ji,et al. Characterization of two copper/zinc superoxide dismutases (Cu/Zn-SODs) from the desert beetle Microdera punctipennis and their activities in protecting E. coli cells against cold[J]. 新疆大学,2019,87:15-27.
APA	Xikeranmu, Zilajiguli,Abdunasir, Maimaitiaili,Ma, Ji,Tusong, Kuerban,&Liu, Xiaoning.(2019).Characterization of two copper/zinc superoxide dismutases (Cu/Zn-SODs) from the desert beetle Microdera punctipennis and their activities in protecting E. coli cells against cold.CRYOBIOLOGY,87,15-27.
MLA	Xikeranmu, Zilajiguli,et al."Characterization of two copper/zinc superoxide dismutases (Cu/Zn-SODs) from the desert beetle Microdera punctipennis and their activities in protecting E. coli cells against cold".CRYOBIOLOGY 87(2019):15-27.