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| 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 |
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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.
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Received: 22 June 2013
Published: 14 August 2013
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