摘要
以甘蔗叶片总RNA为模板,通过RT-PCR扩增细胞色素b6-f复合体铁硫亚基基因的c DNA,采用生物信息学软件分析所克隆基因的编码蛋白特性,并用荧光定量PCR分析该基因的表达情况。结果表明：克隆得到的c DNA片段长度为796 bp,包括1个579 bp的开放阅读框,编码192个氨基酸。甘蔗与高粱CYT基因c DNA序列的同源性为94.0%,氨基酸序列同源性为99.0%;该基因推导的蛋白分子量大小为20.67KD,等电点为6.24,疏水性分值在0.50~2.11;蛋白二级结构中α-螺旋占17.19%,随机卷曲占53.12%,延伸链占21.53%,β-转角只占8.33%,Gen Bank登录号为JQ712582。实时荧光定量分析结果表明,随着甘蔗干旱胁迫时间的延长,So CYT基因表达均呈下降的趋势,PEG与PEG加Si处理的So CYT基因的表达量有显著差异,加硅可减缓So CYT基因表达的下降速度和对细胞色素b6/f复合体的结构的损伤和影响。
So CYT c DNA was amplified using RT-PCR from sugarcane leaves,the characteristics of the deduced protein were analyzed using bioinformatics software and its expression was analyzed using quantitative real-time PCR. The results showed that the sequence contained a796 bp full length fragment with a 579 bp open reading frame,and it encoded a putative So CYT protein with 192 amino acids. Sequences alignment showed that the homology of c DNA sequence between sugarcane and Sorghum bicolor So CYT gene was 94 %,while the homology of amino acid sequence between them was 99 %. The relative protein molecular weight was 20. 67 k D,the PI was 6. 24,and the hydrophobic value was from-0. 5 to 2. 11. The molecular structure prediction results suggested that the protein contained 17. 19 % α-helix,53. 12% loop,21. 53 % extended strand and 8. 33 % β-meander. The sequence has been submitted to Gen Bank with an accession number of JQ712582. The results of quantitative real-time PCR analysis showed that the mRNA of So CYT was decreased with time of PEG stress,significant differences were observed between the treatments of PEG and PEG ＋ Si at 18,26,40,64,96,144 h of the treatment. Silicon can slow down the damage and effect of drought on the structure of the cytochrome b6 / f complex.
出处
《西南农业学报》
CSCD
北大核心
2016年第5期1032-1037,共6页
Southwest China Journal of Agricultural Sciences
基金
国家高技术研究计划(863计划)项目(2013AA102604)
科技部国际合作项目(2013DFA31600)
广西自然科学基金项目(2014GXNSFBA118139)
广西农业科学院基本业务专项项目(桂农2014YD13)
