Construction of Bait Vectors Containing Sucrose Transporter Genes of Hevea Brasiliensis in SOS Recruitment System and Identification of their Autonomous Activation Effects

Key words: 
• Hevea brasiliensis /
• Sucrose transporter /
• bait vector /
• Sos recruitment system /
• yeast two-hybrid system

Abstract: In order to screen the interacting proteins of sucrose transporters （SUTs） from Hevea brasiliensis,the bait vectors of the six sucrose transporters of cytoplasmic yeast two-hybrid system,which was based on Sos recruitment system （SRS）,were constructed and detected for their self activity. The coding sequences of SUTs were obtained by PCR from the plasmids containing respective Hevea SUTs,and the amplified products were cloned into the bait vector pSos at site Sal Ⅰ and Not Ⅰ. Restriction analysis and sequencing confirmed the sequence identity and the correct in frame fusion with the Sos gene of pSos. The resultant bait vectors of the 6 HbSUT genes were termed as pSos-SUTs. The pSos-SUTs were transformed into the temperature-sensitive strain cdc25H,and detected for their self activating effects. The results showed that except for pSos-SUT5,the other five bait vectors had no autonomous activation upon the yeast strain,which were suitable for the follow-up studies.