Construction of RNAi vectors for SmNAC1 transcription factors of Salvia miltiorrhiza using Gateway cloning technology.
- Author:
Rong ZHAO
;
Qi-Xian RONG
;
Yu-Zhong LIU
;
Ye SHEN
;
Lu-Qi HUANG
- Publication Type:Journal Article
- MeSH:
Cloning, Molecular;
methods;
Genetic Vectors;
genetics;
Plant Proteins;
genetics;
Polymerase Chain Reaction;
RNA Interference;
Reproducibility of Results;
Salvia miltiorrhiza;
genetics;
Transcription Factors;
genetics
- From:
China Journal of Chinese Materia Medica
2014;39(9):1569-1573
- CountryChina
- Language:Chinese
-
Abstract:
NAC transcription factors involved in plant growth and development, as well as responses to biotic and abiotic stress. RNAi Vectors for SmNAC transcription factors of Salvia miltiorrhiza was constructed by using Gateway cloning technology, in order to further study the function of SmNAC1 transcription factor. According to Gateway cloning technology, the specific fragments of SmNAC1 containing attB adapter was amplified by PCR using ultra-fideling phusion polymerase of NEB. By the BP recombination reaction, the PCR product containing attB was transferred to an donor vector (pENTR/SD/D-TOPO). Finally, SmNACi specific gene was cloned into pK7GWIWG2D plant expression vectors by LR recombination reaction. Experimental results showed that Gateway cloning technology provide a rapid and highly efficient way to clone the interested gene.
