Construction of plant expression vectors with PMI gene as selection marker and their utilization in transformation of Salvia miltiorrhiza f. alba / 中国中药杂志
China Journal of Chinese Materia Medica
;
(24): 1209-1213, 2014.
Article
in Chinese
| WPRIM
| ID: wpr-321337
ABSTRACT
<p><b>OBJECTIVE</b>To construct plant expression pCAMBIA1301-PMI by substituting PMI for hygromycin resistance gene in pCAMBIA1301 and obtain transgenic Salvia miltiorrhiza f. alba using PMI-mannose selection system.</p><p><b>METHOD</b>The 6-phosphomannose isomerase gene (PMI) of Escherichia coli was amplified by PCR. Sequence analysis showed that it shared 100% amino acids identities with the sequences of PMI genes isolates reported in the NCBI. Based on pCAMBIA1305, the plant expression pCAMBIA1305-PMI was constructed successfully by substituting PMI for hygromycin resistance gene in pCAMBIA1305. pCAMBIA1305-PMI was transformed into Agrobacterium tumefaciens LBA4404, and then the leaves of S. miltiorrhiza f. alba were inoculated in LBA4404 with pCAMBIA1305-PMI.</p><p><b>RESULT</b>Plant expression pCAMBIA1301-PMI was successfully constructed and the leaves of S. miltiorrhiza f. alba inoculated in LBA4404 with pCAMBIA1305-PMI were selected on medium supplemented with a combination of 20 g x L(-1) mannose and 10 g x L(-1) sucrose as a carbon source. The transformation efficiency rate was 23.7%.</p><p><b>CONCLUSION</b>Genetic transformation was confirmed by PCR, indicating that a new method for obtaining transgenic S. miltiorrhiza f. alba plants was developed using PMI-mannose selection system.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Pharmacology
/
Transformation, Genetic
/
Hygromycin B
/
Biomarkers
/
Gene Expression
/
Cinnamates
/
Plants, Genetically Modified
/
Salvia miltiorrhiza
/
Escherichia coli Proteins
/
Escherichia coli
Language:
Chinese
Journal:
China Journal of Chinese Materia Medica
Year:
2014
Type:
Article
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