Cloning and subcellular localization of the GHWAT1-35 gene in Gossypium hirsutum
[Objective]To explore the role of GHWAT1-35 gene in the development of cotton fibers.[Methods]In this study,the fiber at different developmental stages post-anthesis of Gossypium hirsutum va-riety Xi9 were used as materials.The full-length cDNA sequence of the GHWAT1-35 gene was successfully cloned and subjected to bioinformatics analysis,real-time fluorescence quantitative(qRT-PCR)analysis,and subcellular localization.[Results]The length of the GHWAT1-35 gene was 1125 bp,encoding 374 ami-no acids,with a relative molecular weight is 40.23 kDa and a theoretical isoelectric point of 8.74.Protein mul-tiple sequence alignment and construction of a systematic evolutionary tree analysis showed that the GHWAT1-35 protein was most closely related to the Gossypium arboreum.The expression of the GHWAT1-35 gene signif-icantly increased at 15 days after flowering,and subcellular localization predicted that the protein was located on the cytoplasmic membrane.The GHWAT1-35 gene was recombined with the pCAMIA1300-35S-YFP vector to construct a fusion expression vector,which was then transformed into Agrobacterium using the freeze-thaw method and after being injected into tobacco.The GHWAT1-35 protein was observed to be located on the cytoplasmic membrane.[Conclusion]The GHWAT1-35 gene plays an important role in fiber develop-ment,providing a foundation for further exploration of its biological function in Gossypium hirsutum.
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