摘要
为了从雨生红球藻中获得感知蓝光信号的植物类型隐花色素(CRY)序列,采用同源克隆和RACE相结合的方法获得了雨生红球藻编码植物类型Hae-P-CRY的cDNA全长,并对其序列进行生物信息学分析。结果表明,Hae-P-CRY基因的cDNA全长为3608 bp,包含开放阅读框全长2988 bp,编码995个氨基酸,预测等电点6.19,理论分子质量为107.7 ku。经Blast P分析发现,Hae-P-CRY氨基酸序列与莱茵衣藻来源植物类型CreCRY氨基酸序列的相似性达到66.6%,与拟南芥CRY氨基酸序列相似性为46.94%。系统进化分析表明,Hae-P-CRY与其他真核绿藻来源的植物型CRY聚在一支,属于植物类型CRY。结构域分析表明,Hae-P-CRY基因含有光感知PHR结构域和信号转导CCT结构域,暗示具有感知和转导光信号功能。试验从雨生红球藻得到植物类型Hae-P-CRY基因序列,可为其表达、功能及互作蛋白研究奠定基础,同时为进一步解析雨生红球藻在蓝光响应中的分子机制奠定基础。
In this article,a full-length complementary DNA(cDNA)sequence of plant type CRY(designated Hae-P-CRY)was cloned from the green alga Haematococcus pluvialis.The homology cloning and rapid-amplification of cDNA ends(RACEs)methods were applied to get the sequence of Hae-P-CRY.The result showed that the cDNA sequence length of Hae-P-CRY gene was 3608 bp,which contained 2988 bp open reading frame,294 bp 5′-untranslated region(UTR),and 198 bp 3′-UTR with the characteristic of the poly(A)tail.The deduced protein(995 amino acids)had a calculated molecular mass of 107.7 ku with an estimated isoelectric point(pI)of 6.19.According to Blast P analysis,the similarity between Hae-P-CRY amino acid sequence and CreCRY amino acid sequence of Chlamydomonas reinhardtii was 66.6%,and that between Hae-P-CRY and CRY amino acid sequence of Arabidopsis was 46.94%.Phylogenetic analysis showed that Hae-P-CRY was a plant type CRY,which was clustered together with other eukaryotic green algae derived CRY.The results of domain analysis showed that Hae-P-CRY gene contained light sensing PHR domain and signal transduction CCT domain,suggesting that Hae-P-CRY gene had the function of sensing and transmitting light signals.The gene sequence of Hae-P-CRY from Haematococcus pluvialis can lay a foundation for the study of its expression,function and interaction protein,as well as for the further analysis of the molecular mechanism of Haematococcus pluvialis in blue-light response.
作者
杭伟
张宏江
马浩天
王晓丹
许文鑫
赵春超
李润植
崔红利
HANG Wei;ZHANG Hongjiang;MA Haotian;WANG Xiaodan;XU Wenxin;ZHAO Chunchao;LI Runzhi;CUI Hongli(Institute of Molecular Agriculture and Bioenergy,Shanxi Agricultural University,Taigu 030801,China)
出处
《山西农业科学》
2020年第5期677-682,共6页
Journal of Shanxi Agricultural Sciences
基金
国家自然科学基金项目(31902394)
山西省应用基础研究项目(201801D22125)
辽宁省科学技术计划项目(20170540047)
山西省煤基重大科技专项(FT-2014-01)
山西省重点研发重点项目(201603D312005)
山西农业大学科技创新基金项目(2018YJ16)
山西省研究生教育创新项目(2019SY226)。
关键词
雨生红球藻
隐花色素
基因克隆
生物信息学分析
Haematococcus pluvialis
cryptochrome
gene cloning
bioinformatics analysis
