摘要
通过同源克隆从木薯品种辐选01(RS01)中获得SPS基因保守序列,利用RACE扩增技术获得全长cDNA序列并使用相关软件进行生物信息学分析,并采用实时荧光定量PCR分析木薯SPS基因在不同时期、不同组织中的相对表达量。结果表明:克隆获得的木薯SPS基因cDNA序列全长3 857bp,开放阅读框(ORF)长3 228bp,编码1 076个氨基酸,命名为MeSPS(GenBank登录号KX822780);同源性分析表明,MeSPS基因与麻风树、蓖麻和荔枝的SPS基因序列同源性较高,分别为89%、89%和87%;其氨基酸序列与其他植物SPS氨基酸同源性在77%-92%;经qRT-PCR分析结果表明,MeSPS在苗期的相对表达量较高;在同一生长时期,MeSPS在叶片中相对表达量高于茎段和块根。
Sucrose phosphate synthase(SPS)gene from cassava RS01 was cloned to study its biological functions.The conserved fragment of the gene was first obtained by homologous cloning,and its full length cDNA by rapid amplification of cDNA ends(RACE).The characteristics,gene sequence and other biological information retrieved were analyzed.Expressions of the genes from various tissues of the plants at different times were compared by RTqPCR.The results showed that the cDNA of SPS gene had a full length of 3,857 bp,and an ORF of 3,228 bp encoding aprotein of 1,076 amino acids.It was subsequently named MeSPS with a GenBank number of KX822780.Its homologous alignments with the nucleotide and protein databases were highly homologs with those of Jatropha carcas L.,Ricinus communis L.,and Litchi chinensis Sonn.at 89%,89%,and 87%,respectively.Its amino acid sequence was 77%-92% in similarity with the SPS genes of other plants.The qRT-PCR analysis showed that MeSPS gene was highly expressed during seeding,and,was more expressed in the leaves than the stems or roots on aplant in a same growth period.
出处
《福建农业学报》
CAS
北大核心
2016年第12期1273-1279,共7页
Fujian Journal of Agricultural Sciences
基金
国家"973"计划项目(2010CB126601)
广西科学研究与技术开发计划项目(桂科重14121005-2-1)
南宁市科学研究与开发计划项目(201109044B)
国家现代农业产业体系广西薯类产业创新团队项目(nycytxgxcxtd-03-11-2)
南宁市科学研究与技术开发计划项目(20132307)
关键词
木薯
蔗糖磷酸合成酶
基因克隆
表达分析
cassava
sucrose phosphate synthase
gene cloning
expression analysis