The Cloning and Haplotype Analysis of MAP30 Gene from Bitter Gourd

Purpose The aim of this study was to reveal the complete gene structure and haplotypes of MAP30 in bitter gourd, expected to provide reference for studying the regulation mechanism of MAP30 gene expression.

Method Taking the primary core collection of bitter gourd as material, the MAP30 gene was cloned through referring to the genome sequence of bitter gourd, the complete gene structure and haplotypes of MAP30 were analyzed through adopting bioinformatic softwares.

Results The MAP30 gene was 920 bp in length, including 52 bp 5'-UTR, 7 bp 3'-UTR and 861 bp ORF, with no intron, encoding 286 amino acids. Phylogenetic analysis showed that the MAP30 protein was far from the RIPs protein of other plants of Cucurbitaceae family, so the MAP30 protein was highly conserved in the momordica genus. 6 SNP loci were found through comparing the MAP30 gene sequences of 34 bitter gourd germplasm, the 1^st SNP was located in the region of the 5'-UTR, and the remaining 5 SNPs were located in the coding region. There were 3 haplotypes of MAP30 gene in 34 bitter gourd germplasm through SNP grouping. Three amino acid mutations were found according to comparing the amino acid sequences encoded by the 3 haplotypes. They were M/V in the 2^nd loci, R/H in the 69^th, and N/D in the 74^th, respectively, which were corresponding to the 2^nd, 3^rd and 4^th SNP loci, the 5^th and 6^th SNP loci did not cause amino acid changes.

Conclusion The complete gene structure of MAP30 had been cloned, and the relationship between MAP30 protein and RIPs of other cucurbit plants had been analyzed. In addition, the SNPs distribution and haplotype types of MAP30 gene in different bitter gourd germplasm had also been analyzed.