[Cloning of Scutellaria viscidula CHS gene and construction of its sense and antisense plant expression vectors].
OBJECTIVE
To get Scutellaria viscidula CHS gene sense and antisense plant expression vectors,and explore the mechanism of CHS's effect on the flavonoid biosynthetic pathway through the transformation of Scutellaria viscidula.


METHODS
CHS gene fragment was amplified by Reverse Transcription Polymerase Chain Reaction (RT-PCR) from total RNA of Scutellaria viscidula and cloned into PMD18-T vector, sequenced and bioinformaticly analyzed.


RESULTS
The DNA sequence was correct and we simultaneously guessed the amino acid sequence and the three-dimensional structure of CHS. Cloned fragments and pCAMBIA1304+ vector were digested using Blg II and BstE II at the same time, then they were connected with CHS gene fragment and pCAMBIAI304+ big skeleton fragment to get CHS sense and antisense plant expression vectors.


CONCLUSIONS
The successful construction of Scutellaria viscidula CHS gene sense and antisense plant expression vectors, and the transformation into C58C1 and LBA4404 through the freeze-thaw method, lay the foundation for Agrobacterium-mediated transformation of Scutellaria viscidula CHS gene in plant heredity.
Rao
Can
C
Key Laboratory of Eco-environments, Three Gorges Reservoir Region, Ministry of Education, College of Life Science, Southwest University, Chongqing 400715, China. raocan@swu.edu.cn
Lei
Wei
W
Li
Peng
P
Sun
Yi-ming
YM
Sun
Min
M
D000217:Acyltransferases / Q000737:chemistry / Q000235:genetics*; D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D017931:DNA Primers; D018744:DNA, Plant / Q000235:genetics; D017343:Genes, Plant; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D011995:Recombination, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction; D031363:Scutellaria / Q000201:enzymology* / Q000235:genetics*; D017422:Sequence Analysis, DNA
D017931:DNA Primers; D018744:DNA, Plant; D000217:Acyltransferases; C020141:flavanone synthetase