Ginkgo Flavonoid Synthase Gene Cloning and Characterization

Based on an mRNA fragment annotated in the RNA-seq library of ginkgo (Wu et al., 2018 (link)), nested primers were designed to amplify full-length cDNA using the SMARTer RACE 5’/3’ Kit (Clontech, Japan) (Supplemental Table S1). Open reading frames (ORFs) were predicted by the NCBI ORFfinder, and the ORF of the GbFLSa gene was cloned to further determine its function. Moreover, ExPASy online software, DNAMAN 6.0 software, MEGA 7.0 software, SOPMA software and Gene Structure Display Server online software were used to analyze the FLS protein sequence (Wu et al., 2020a (link); Wu et al. 2020b (link)).

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Guo J., Wu Y., Wang T., Xin Y., Wang G., Zhou Q, & Xu L.A. (2023). GbFLSa overexpression negatively regulates proanthocyanin biosynthesis. Frontiers in Plant Science, 14, 1093656.

Publication 2023

SMARTer RACE 5’/3’ Kit

Cdna Gene Ginkgo Library Mrna Open reading frames Primers Protein sequence Rna seq

Corresponding Organization : University of British Columbia

Other organizations : Zhejiang Academy of Forestry

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Variable analysis

independent variables

MRNA fragment annotated in the RNA-seq library of ginkgo
Nested primers designed to amplify full-length cDNA using the SMARTer RACE 5'/3' Kit
ORF of the GbFLSa gene

dependent variables

Full-length cDNA
Function of the GbFLSa gene

control variables

Not explicitly mentioned

controls

No positive or negative controls were explicitly mentioned in the provided information.

Annotations

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