Molecular Analysis of the Gene LAS17 Mediating T-DNA Trafficking inside yeast cells

BATHULA HARIPRIYA

Publication

Molecular Analysis of the Gene LAS17 Mediating T-DNA Trafficking inside yeast cells

BATHULA HARIPRIYA

Abstract

Agrobacterium tumefaciens is known for its applications in plant genetic engineering for its unique ability to transfer a segment of its DNA (T-DNA) from its tumor-inducing (Ti) plasmid into plant cells, fungi and mammalian cells. Agrobacteriummediated transformation is the only known case of trans-kingdom DNA transfer that occurs in nature. The ability of Agrobacterium tumefaciens to mediate trans-kingdom transfer of genetic material has established an exciting paradigm in the field of genetic manipulation. It has been established that under laboratory conditions, Agrobacterium can also transfer T-DNA into a wide range of other eukaryotic species, including yeast cells. To date, scientists have obtained a comprehensive understanding of Agrobacterium proteins that mediate the transfer process, though the involvement of host proteins remains unclear. The current study aims to use yeast Saccharomyces cerevisiae as a eukaryotic model to identify and characterize host factors involved in Agrobacterium-mediated transformation (AMT). So far, the genetic screening of yeast mutants has revealed that the knock-out of Las17 results in a significant increase in AMT efficiency. In the current study, a series of genetic and bio-imaging approaches have been adopted to study the role of Las17 gene in the T-DNA trafficking inside the yeast cells. The results show that TDNA is trafficked more efficiently in Las17 mutant cells implying that the Agrobacterium mediated transformation process employs an endocytosis-independent pathway.