Overexpression of the novel gene CIA7 in Chlamydomonas reinhardtii (995.3)

Abstract

Heavy metal pollution has become a significant environmental and public health problem that the world faces. C. reinhardtii is an excellent model organism that has often been used in studies of heavy metal tolerance. This alga contains the novel gene CIA7 that possibly encodes a protein that may play a role in metal binding. This gene was isolated from inorganic carbon acquisition mutants. By cloning and overexpressing CIA7 in C. reinhardtii, it can be determined if CIA7 is critical in C. reinhardtii’s metal toxicity tolerance. This gene of interest was first extracted from the alga and then amplified using PCR. Vectors pENTR™/D‐TOPO® (Invitrogen) and psL72 plasmids were used for cloning CIA7 in E.coli. C.reinhardtii cells were then transformed through electroporation. Growth analysis between wild type C.reinhardtii D66 and transformants at different metal concentrations were done to determine the effect of CIA7 on C.reinhardtii under metal stress. If a link between this protein and heavy metal tolerance is identified, this could potentially lay the foundation for its use in heavy metal pollution.Grant Funding Source: Supported by: Norman Hackerman Advanced Research Program