Open AccessIsolation of cadmium sensitive mutants in Chlamydomonas reinhardtii by transformation/insertional mutagenesis
Author(s) -
McHugh Joyce Pfeifer,
Spanier Jonathan G.
Publication year - 1994
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1994.tb07290.x
Subject(s) - chlamydomonas reinhardtii , insertional mutagenesis , mutant , plasmid , mutagenesis , transformation (genetics) , biology , chlamydomonas , genetics , gene , strain (injury) , wild type , cadmium , microbiology and biotechnology , chemistry , organic chemistry , anatomy
An arg7, cw15, mt + strain of Chlamydomonas reinhardtii (CC1618) was transformed with pARG7.8, a plasmid containing the wild‐type ARG7 gene. Over 2300 arg + transformants were selected on TAP media. Upon subsequent analysis on TAP plus cadmium plates, five of the transformants failed to grow at a level of 400 μM cadmium and were designated as cadmium sensitive (Cd s ) mutants. Hybridization data indicated that vector (pBR329) sequences were present in these five mutants, but not in the untransformed parental strain. Two of the mutants have been back crossed to an arg7, cw15, Cd + , mt − strain (CC425) and found to have progeny which always cosegregate the arg + and Cd s phenotypesin these two mutants results from the insertion of the plasmid pARG7.8 into a gene involving cadmium detoxification, and it provides a method by which to clone the interrupted gene(s).