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采用电子克隆与RT-PCR相结合的方法获得茶树硫酸盐转运蛋白(CsSUL3.5)的cDNA序列,并进行了相关生物信息学分析。克隆到茶树CsSUL3.5 cDNA序列2 017 bp(GenBank登录号KP984500),包含完整的开放阅读框(ORF)1914 bp,编码637个氨基酸。生物信息学分析显示,CsSUL3.5编码的蛋白分子量为70.38 kD,无信号肽,是疏水性的非分泌蛋白,有11个跨膜区;与其他物种相似性均在60%以上,与烟草的相似性最高(74%),具有硫酸盐转运蛋白家族典型的保守结构域和空间结构,属于硫酸盐转运蛋白家族。系统进化分析表明茶树的CsSUL3.5属于第3亚家族,与芝麻的关系比较近。荧光定量PCR表明该基因在茶树幼苗根与成熟叶中均有表达;Na_2SO_4与Na_2SeO_4处理均能显著诱导CsSUL3.5的表达。
The cDNA sequence of tea sulfate transporter (CsSUL3.5) was obtained by a combination of electronic cloning and RT-PCR, and the related bioinformatics analysis was carried out. The full-length open reading frame (ORF) of 1914 bp, encoding 637 amino acids, was cloned into the cDNA library of tea tree CsSUL3.5 with a length of 2017 bp (GenBank accession number KP984500). Bioinformatics analysis showed that the protein encoded by CsSUL3.5 had a molecular weight of 70.38 kD and no signal peptide. It was a hydrophobic non-secreted protein with 11 transmembrane domains. The similarity to other species was over 60% The highest similarity (74%), which has a typical conserved domain and spatial structure of the sulfate transporter family, belongs to the sulfate transporter family. Phylogenetic analysis showed that CsSUL3.5 belongs to the 3rd subfamily of tea plant, and has a close relationship with sesame. Quantitative real-time PCR showed that the gene was expressed in both seedling and mature leaves of tea plant, and both Na_2SO_4 and Na_2SeO_4 significantly induced the expression of CsSUL3.5.