摘要
从茶树叶片中提取总RNA ,经反转录合成cDNA第一链。以cDNA第一链为模板 ,分别用三对PCR引物扩增SAM合成酶基因的中间主片段、 3 端和 5 端片段 ,最后经BLAST比较及重叠区域拼接得到完整的SAM基因序列。所得序列长 130 3bp ,编码 394个氨基酸。与中华猕猴桃、番茄、长春花SAM合成酶基因的核苷酸序列的同源性分别为 87% ,83 %和 83%。氨基酸序列与三角杨、猕猴桃、番茄、长春花等的SAM氨基酸序列的同源性为 92 %~ 90 %。证实所获得的基因序列为茶树SAM合成酶基因。该基因的克隆为今后进一步开展茶树逆境生理。
Total RNA was extracted from tea leaf and the first strand of cDNA was synthesized with reverse transcription The principal fragment,3' end fragment and 5' end fragment of S adenosylmethionine synthetase gene (SAM gene)was amplified with the first strand cDNA as template and three pairs primers by PCR Complete SAM gene sequence was obtained by BLAST comparison of the three fragments and splicing according to the overlapping regions The sequence of the SAM gene is 1303 base pairs (bp)in length with an opening reading frame (ORF) encoding 394 amino acids The cDNA sequence shows a significant homology to the SAM genes from Actinidia chinensis (87%),Lycopersicon esculentum (83%),Catharanthus roseus (83%) The amino acid sequences shares 90%-92% identity with SAM synthetase from Populus trichocarpa,Actinidia chinensis,Petunia hybrida,Catharanthus roseus, etc Based on sequence homology,it was suggested that the obtained cDNA was a SAM gene in tea plant The isolation of the SAM gene established a good foundation for further study on stress physiology,senescence physiology and caffeine biosynthesis in tea plant
出处
《茶叶科学》
CAS
CSCD
北大核心
2001年第1期21-25,共5页
Journal of Tea Science
基金
中 -日国际合作项目!"C0 0 5 6"研究内容之一
关键词
茶树
SAM合成酶
基因克隆
RT-PCR
Camellia sinensis
S adenosylmethionine synthetase
gene cloning
RT PCR