Cloning and CRISPR/Cas9 gene editing vector construction of McPDS in bitter gourd(Momordica charantia L.)
HAN Xin
GUO Jinju
ZHANG Huiyao
WU Tingquan
ZHANG Changyuan
Abstract：A gRNA-specific CRISPR/Cas9 gene editing vector targeting phytoene dehydrogenase(PDS)gene of bitter gourd was constructed,hoping to lay a foundation for the establishment of CRISPR/Cas9 gene editing technology system of bitter gourd.In this study,the CDS region of McPDS gene was cloned from the leaf cDNA of bitter gourd inbred line B07.The results showed that the coding region length of McPDS was 1731 bp,encoded 576 amino acids,the theoretical relative molecular weight was 64.44 kD,and the theoretical isoelectric point(PI)was 7.09.The transmembrane structure analysis showed that the protein was a hydrophilic non-transmembrane protein.Phylogenetic analysis showed that McPDS had high homology with PDS proteins in cucurbit plants such as cucumber and melon.In addition,using McPDS as the target gene,two highly specific targets were screened at the 5'end,primers were designed,and a double-target CRISPR/Cas9 gene editing vector was successfully constructed,which laid a technical foundation for the establishment of bitter gourd gene editing system.
Keywords：Bitter gourdMcPDSGene cloningVector constructionGene editing
Publication Date：2024-03-05
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：8( 20-27 )