SITE-DIRECTED MUTAGENESIS IN THE ACTIVE-SITE OF THE HERPES-SIMPLEX VIRUS TYPE-1 THYMIDINE KINASE GENE

被引:8
作者
FETZER, J [1 ]
FOLKERS, G [1 ]
MULLER, I [1 ]
KEIL, GM [1 ]
机构
[1] FED RES CTR VIRUS DIS ANIM, TUBINGEN, GERMANY
关键词
HSV-1 THYMIDINE KINASE; SITE-DIRECTED MUTAGENESIS; HSV-1 TK ANTISERUM;
D O I
10.1007/BF01702400
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
The thymidine kinase (TK) of herpes simplex virus type 1 (HSV-1) contains three regions of homology to other ATP utilizing enzymes. We have altered one region of the protein, which seems to play an important role in phosphorylating substrates by site-directed mutagenesis. When the aspartate 162 was changed to asparagine, the enzyme lost its activity. To identify the inactive protein, expressed by a vaccinia vector in eukaryotic cells, a monospecific antiserum against a bacterial tryptophan E-HSV-1 TK fusion protein was made. These results support the suggestion that aspartate 162 is essential for the enzymatic activity.
引用
收藏
页码:205 / 209
页数:5
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