目的 克隆刺五加的液泡膜内在蛋白（tonoplast intrinsic proteins，TIP）基因，并对其进行生物信息学和表达分析。方法 采用cDNA末端快速扩增（RACE）技术克隆刺五加TIP基因cDNA的全长序列。以GAPDH为内参照基因，通过RT-PCR法检测TIP基因在不同生长发育时期和器官中的表达情况。结果 刺五加TIP基因cDNA的全长1 080 bp，开放阅读框长756 bp，编码251个氨基酸的蛋白，该蛋白包含TIP家族的标志性序列。刺五加的TIP蛋白具有6个跨膜螺旋，定位于液泡膜。表达分析结果显示，刺五加TIP基因在不同生长发育时期和不同器官中均有表达，但表达量具有显著差异（P＜0.05）。其中盛花期的表达量最高，是最低量果实快速生长期的2.07倍；各器官中，叶片的表达量最高，是最低量根的1.73倍。结论 首次分离到刺五加TIP基因的cDNA全长序列，并证实其在盛花期的叶中表达量最高，为进一步研究TIP基因对刺五加水分代谢的影响奠定基础。

Objective To clone tonoplast intrinsic proteins (TIP) gene from Eleutherococcus senticosus and to analyze its bioinformatics and expression. Methods Full length cDNA of TIP gene was cloned by rapid amplification of cDNA ends (RACE). Taking GAPDH gene as reference gene, the expression of TIP in different organs of E. senticosus at various growing periods was detected by RT-PCR. Results The full length cDNA of TIP gene was 1 080 bp containing a 756 bp open reading frame (ORF) that encoded a protein of 251 amino acids including the typical sequences of TIP family. TIP gene was located in tonoplast with six transmembrane domains. The result of expression analysis indicated that TIP gene expressed in different growth periods and organs of E. senticosus, and the expression amount differed significantly (P < 0.05). The highest content of the expression showed up at full opening flower stage, which was 2.07 times as much as that in the lowest at rapid fruit growth stage. The highest content of the expression was in the leaves which was 1.73 times as much as that of the lowest in roots. Conclusion We have first extracted the full length cDNA of TIP gene in E. senticosus, which proves that the highest content of the expression is in the leaves at full opening flower stage. This work provides a foundation for the fuither investigation on the ffect of water metabolism in E. senticosus.

河北省自然科学基金——石药集团医药联合研究基金项目（H2012401006）；河北联合大学培育基金（GP201306）