ISSN:
1572-9931
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
,
Medicine
Notes:
Abstract The complete hamsterERCC2 cDNA was constructed in a plasmid vector from clones of three overlapping reverse transcribed/polymerase chain reaction amplified fragments using unique restriction enzyme recognition sites within the regions of overlap. This complete cDNA insert was then cloned into a mammalian expression vector, pcD2E, and tested for function by the ability to confer UV resistance to theERCC2 mutant CHO cell line UV5. Site-specific mutagenesis was used to introduce the G347→A and G1844→A changes resulting in the Cys 116→Tyr and Gly615→Glu mutations previously identified in UV5 and UVL-13 (also anERCC2 mutant CHO cell line), respectively. The 116Tyr and 615Glu plasmids each failed to confer UV resistance to UV5 or UVL-13 cells, respectively, demonstrating that the changes identified are indeed the causative mutations in UV5 and UVL-13.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF02369437
