Study On Cpcp (Pollen Coat Protein Gene) Promoter Expression Of Tea Plant (Camellia Sinensis)

The promoter of tissue-specific expression usually have the particular regulatory elements, which make gene just carried on the expression in the particular tissue and organs and the particular growth stages by working with trans-acting factor. With the rapid development of transgenic technologies application, fast isolation and authenticate various tissue-specific expression promoter of plant has already become the hot point and difficulty on the plant genetic engineering research.It will take about one year and a half from bud Formation to seed maturation of tea plant(Camellia sinensis), thus consumed a great deal of nutrient.Therefore, controling reproductive growth and promoting vegetative growth is a key that raises the tea yield and qualities.In the process of pollen development, there are various tissue-specific expression promoters. In this work, we constructed a eukaryotic expression vector of Cpcp promoter, and Agrobacterium-mediated genetic transformation is used to transfer Cpcp into different tissue and organs of different plant.Then we analyzed the expressing activity of Cpcp promoter via expression of gus reporter gene in different tissue and organs of different plant. It could provide some research foundation on controling reproductive growth and promoting vegetative growth in order to raises the tea yield and qualities or Male Sterility etc by use of the specific expression of Cpcp promoter. The results were as follows:(1)The Cpcp promoter were PCR amplified with one pairs of primers based on the sequences in GenBank (Accession no. EF116920, EU255276).Then we constructed a eukaryotic expression vector of Cpcp promoter pBI-Cpcp,and transfer it into Agrobacteriurn tumefaciens.(2) We analyzed expression of gus reporter gene in different tissue and organs of different plant by GUS staining and fluorescence quantitatiy,which in response to expressing activity of Cpcp promoter.and the result indicate that the GUSgene was driven by Cpcp promoter and is strongly expressing in pollen.(3)The eukaryotic expression vector of Cpcp promoter was transformed into tobacco with leaf disc method mediated by Agrobacterium tumefaciens. The Cpcp promoter fragment were PCR amplified with one pairs of primers based on the Cpcp sequences.Therefor we had obtainedsome transgenic tobacco.