The invention discloses a
Carya cathayensis auxin efflux carrier protein CcPILS
gene cloning and
expression analysis method. The method includes the steps of: material preparation,
RNA extraction and purification, CcPILS
gene RACE,
bioinformatics analysis, and temporal and spatial
expression analysis. The method provided by the invention clones
Carya cathayensis PIL
homologous gene CcPILS by RACE technique, the
gene has a full-length sequence of 1541bp, wherein the
open reading frame sequence is 1263bp,
bioinformatics software analysis is applied to predict that the sequence encodes 420 amino acids, the CcPILS
protein molecular weight is about 46.22KD, PI is 5.38, the
protein is located at an
endoplasmic reticulum membrane, is equipped with 5 transmembrane hydrophobic structural domains respectively at the N terminal and C terminal, and is separated by a middle hydrophilic loop. The gene has
high homology with
Arabidopsis thaliana AtPILS5 and AtPILS7, and belongs to
Clade III subtribe of PILS
gene family.
Fluorescence quantitative RT-
PCR analysis shows that before and after
Carya cathayensis grafting, CcPILS has expression trend in scion consistent with that in
rootstock, and provides a beneficial promoting effect on solution of the problems of difficult Carya cathayensis
grafting, difficult variety breeding and difficult quality improvement, etc.