Moclecular Cloning and Sequence Analysis of SPT Gene Transcription Factor
Regulation of Pistil from Cabbage
YANG Pu-li，XU Jun-qiang，LIU Zhi-yu，WANG Zhi-min，ZHANG Dan-hua，TANG Qing-lin*，
SONG Ming*
College of Horticulture and Landscape Architecture， Southwest University， Key Laboratory of Horticulture Science for
Southern Mountainous Regions， Ministry of Education， Key Laboratory of Olericulture， Chongqing 400715， China
Abstract：Taking cabbage（Brassica oleracea L.） E1 as materials，We extracted total RNA
from capullo，reverse transcribed cDNA. According to SPT gene sequence of Arabidopsis，primers
were designed and 1 085 bp SPT gene with 1 062 bp open reading frame（ORF）was cloned by homology
cloning techniques. The deduced BoSPT protein contained 353 amino acids，with a molecular
weight of 37.67 kD and pI of 6.83. After double enzyme of EcoRI and KpnI restriction enzymes，
and then construct the recombinant plasmids pET43.1a-BoSPT. After transformation to E. coli Rosetta（
DE3），the expression of recombinant proteins were detected via SDS-PAGE. The structural
analysis of BoSPT though Smart-embl showed that it contained bHLH family domain，which located
at the position of 173-221 amino acid residues，The phylogenetic tree indicated that the BoSPT had close genetic relationship with AtSPT and AlSPT. Prokaryotic expression showed that the molecular
mass of BoSPT protein was purified.
