Green Alga (Chlamydomonas reinhardtii).
This protocol describes the Agrobacterium tumefaciens-mediated nuclear transformation of a microalgae Chlamydomonas reinhardtii, using a gene construct carrying the genes coding for beta-glucuronidase (gus), green fluorescent protein (gfp), and hygromycin phosphotransferase (hpt). The transformation frequency with this protocol as revealed by hygromycin resistance was many fold higher (about 50-fold) than that of the commonly used glass bead method of transformation. The simplicity of Agrobacterium-mediated gene transfer and the high transformation frequency as well as the precision of T-DNA integration will enable further molecular dissection of this important model organism as well as other algal systems to understand basic plant metabolic processes as well as to exploit the systems for biotechnological applications.
Rajam
Manchikatla V
MV
Plant Polyamine and Transgenic Research Laboratory, Department of Genetics, University of Delhi - South Campus, Benito Juarez Road, New Delhi 110 021, India.
Kumar
S Vinod
SV
D016960:Agrobacterium tumefaciens / Q000166:cytology / Q000235:genetics*; D000818:Animals; D018929:Cell Culture Techniques; D016825:Chlamydomonas reinhardtii / Q000166:cytology / Q000235:genetics* / Q000502:physiology; D018920:Coculture Techniques; D003470:Culture Media; D048028:DNA, Algal / Q000737:chemistry; D004269:DNA, Bacterial; D018014:Gene Transfer Techniques; D005966:Glucuronidase / Q000032:analysis; D049452:Green Fluorescent Proteins / Q000032:analysis; D014170:Transformation, Genetic; D019076:Transgenes
D003470:Culture Media; D048028:DNA, Algal; D004269:DNA, Bacterial; C036483:T-DNA; D049452:Green Fluorescent Proteins; D005966:Glucuronidase