Cloning,Expression And Regulation Of HbPEPRK1 Gene From Hevea Brasiliensis

Ca2+ signal is widely involved in physiological process in plant,such as plant growth and development,metabolic regulation,hormone and stress responses which maintain normal growth environment for plant.Ca2+ sensor proteins are not only able to perceive Ca2+ signal but also transmit it.Phospnoenolpyruvate carboxylase kinase-related kinase(PEPRKs)are one type of Ca2+ sensor proteins.PEPRKs are one type of Ser/Thr protein kinase and have typical Ser/Thr protein kinases catalytic conservative sequence.PEPRKs and calcium dependent protein kinase(CDPKs)which is not only widely studied but also one type of Ca2+ sensor responder are highly homologous.PEPRKs are only exist in plants,but the current the knowledge of PEPRKs is less to accumulate.In this study,a PEPRK1 from rubber tree coding phosphoenolpyruvate carboxylase kinase-related kinase was cloned.The biological information,structure characteristics and expression regulation of HbPEPRK1 are studied.The main research results was follows:HbPEPRKl from rubber tree coding phosphoenolpyruvate carboxylase kinase-related kinase was cloned.Sequence analysis revealed that HbPEPRKl cDNA length 1401bp,encoding 466 amino acids,PI 8.88.HbPEPRKl protein shared high identities to phosphoenolpyruvate carboxylase kinase-related kinases from other plants,and had the typical structure of plant Serine/Threonine protein kinase.A 2150 bp promoter regions of HbPEPRKl was isolated from rubber tree,in which the ethylene response elements ERE,defense and stress response TC-rich repeats,drought response element MBS and damage response element WUN-motif were identified.These structure characteristics showed that HbPEPRKl might be involved in the growth of rubber tree and response to environmental stress.The real-time RT-PCR results indicated that HbPEPRKl was expressed in all detected organs,in which,the transcripts of HbPEPRK1 was higher in latex than in other organs.The expression of HbPEPRKl was significantly induced in latex by ethephon.Prokaryotic expression vector pGEX-HbPEPRK1 was constructed and HbPEPRKl target protein is successful to express in the E.coli Transetta(DE3).Fifteen interaction proteins involved in protein synthesis,protein modification and activity regulation,rubber biosynthesis,and signal transduction of HbPEPRKl were screened by yeast two-hybrid.