生物技术通报2023，Vol.39Issue(11)：318-327,10.DOI:10.13560/j.cnki.biotech.bull.1985.2023-0551
钩吻脂氧合酶基因GeLOX1的鉴定及低温胁迫表达分析
Identification of the Lipoxygenase Gene GeLOX1 and Expression Analysis Under Low Temperature Stress in Gelsmium elegans
摘要
Abstract
In recent years,the medicinal and feeding value of Gelsemium elegans has become increasingly prominent.However,G.elegans is not tolerant to low temperatures during its growth process.Exploring its low temperature response genes may lay the foundation for the research of cold resistance breeding of G.elegans.Lipoxygenase(LOX)has a significant impact on the physiological and biochemical processes of plants in seed aging,stress resistance,and other aspects.The LOX genes of G.elegans in response to low temperature stress were mined from the transcriptome database of G.elegans constructed by our team.A full-length cDNA sequence of the GeLOX1 gene was cloned from the leaves of G.elegans using RT-PCR technology,and its bioinformatics,subcellular localization,gene expression,prokaryotic expression,and plate stress were analyzed.The results showed that the amino acid length of the protein encoded by the GeLOX1 was 761 aa,and the relative molecular weight of the protein was 87.00 kD.It was predicted to be an unstable hydrophilic protein,containing 28 serine phosphorylation sites,22 threonine phosphorylation sites,and 9 tyrosine phosphorylation sites.The analysis results of evolutionary tree indicated that GeLOX1 belonged to the 9-LOX family.The subcellular localization test results showed that the GeLOX1 was located in the cytoplasm.Real time fluorescence quantitative PCR analysis revealed that the GeLOX1 was highly expressed in the roots of the G.elegans,and its expressions showed a downward trend under 4℃low temperature stress.After prokaryotic expression induction,the recombinant protein of the GeLOX1 showed a target band at approximately 111 kD,and the accumulation of the recombinant protein reached its peak at 8 h of induction.In addition,the plate stress experiment showed that the prokaryotic expression strain of the GeLOX1 was more sensitive to low temperature stress compared to the control group.The research results indicate that GeLOX1 of G.elegans can respond to low temperature stress.关键词
钩吻/脂氧合酶(LOX)/序列分析/低温胁迫/表达分析Key words
Gelsemium elegans/lipoxygenase(LOX)/sequence analysis/low temperature stress/expression analysis引用本文复制引用
尤垂淮,苏亚春,谢津津,张婷,崔天真,孙欣路,臧守建,武奕凝,孙梦瑶,阙友雄..钩吻脂氧合酶基因GeLOX1的鉴定及低温胁迫表达分析[J].生物技术通报,2023,39(11):318-327,10.基金项目
福建农林大学大学生创新创业训练计划(X202310389110),福建省卫生厅中青年骨干人才培养项目(2020GGB024),福建省科技厅科技创新联合资金项目(2020Y9066) （X202310389110）
