金银花U6启动子的克隆及转录活性分析

doi:10.3976/j.issn.1002-4026.2022.02.002

Abstract

Abstract:

U6 promoter is an important element that drives sgRNA transcription in the construction of the CRISPR/Cas9 system. In this study, three types of U6 promoters were cloned from Huajin 6 Lonicera japonica using polymerase chain reaction(PCR), and five promoters of different lengths were constructed to drive the GUS expression vector. CaMV35S was used in a positive control experiment, and Agrobacterium tumefaciens was used in a negative control experiment. Tobacco protoplasts were transfected through the transient transformation method using A. tumefaciens. After GUS staining, the quantity and efficiency of protoplasts were observed under a microscope. The staining results revealed that the highly efficient transcription promoter Chr01 U6-F1 was screened successfully, which laid a foundation for the gene editing of Lonicera japonica in the later stage.

Key words: Lonicera japonica, U6 promoter, clone, gene editing

CLC Number:

TANG Zhi-qiang,LI Xiao-li,XU Xiao-han,PU Gao-bin. Analysis of the cloning and transcriptional activity of U6 promoter from Lonicera japonica[J].Shandong Science, 2022, 35(2): 11-17.

Figures/Tables 7

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References 23

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引物名称	正向引物序列	反向引物序列
Chr01 U6-F1	AAGCTTGAAACTAATGTGTATAT	GGATCCGCTTAATTTCCCACCC
Chr02 U61-F1	AAGCTTTCAGGTGGGATTAAAAT	GGATCCTGTCTTATTGTTGTGCG
Chr02 U61-F2	AAGCTTGAGTGTTCGTATCAGTT	GGATCCTGTCTTATTGTTGTGCG
Chr02 U62-F1	AAGCTTTATGTTTGAACTTGATT	GGATCCTACATTGTTGTTGTCCG
Chr02 U62-F2	AAGCTTCTCACATGGGAACTCTA	GGATCCTACATTGTTGTTGTCCG

Analysis of the cloning and transcriptional activity of U6 promoter from Lonicera japonica

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