Isolation And Identification Of Leaf-specific Promoters In Rice

Promoters played an important regulatory role at the transcriptional level.In recent years,research workers found the structure of the promoter and its function,And elaborated on the direction of plant promoters in future studies.Rice was an important food crop of mankind,In recent years,people had cultivated new rice varieties by studying many methods,The study of rice-specific promoters had become a recent hotspot.Using tissue-specific promoters,Improve its specific traits,Is of great importance.The main purpose of this research was,The promoter of vascular bundle sheath-specific expression was isolated from rice,For the cultivation of new varieties of rice to provide technology and method support.Specific research content mainly has the following points:1,Screening the rice enhancer capture system,Screening of vascular sphincter-specific lines by GUS staining.The microstructure of the cross section of rice leaves was observed by hand,21 lines with more obvious staining were screened out.2,21 Lines were obtained by hand slicing,Take the leaves to do paraffin section analysis,It was further confirmed that the GUS gene was specifically expressed in the vascular sheath of the above lines.Five strains of GUS gene-specific lines were obtained for subsequent experiments.3,According to the vector sequence containing the GUS gene,Design primers,By TAIL-PCR method,Cloning the vector upstream sequence,The obtained sequences were compared with wild-type Japanese rice in NCBI,Locate the insertion site.4,PCR method to clone the upstream sequence,Construction of plant expression vectors.5,Agrobacterium transformation Arabidopsis thaliana,Collect T0 on behalf of the seed,Re-plating,Transplanting,Detection of the cloned promoter initiated the expression of the GUS gene.