[Engineered glutamic acid decarboxylase fusion protein in diagnosis of type I diabetes mellitus].
OBJECTIVE
To get recombinant GAD 65 proteins which have immunogenicity by the method of gene engineering so that the recombinant protein can be used for early diagnosis in type I diabetes mellitus.


METHODS
We amplified the complete fragment encoding GAD65 gene by PCR, and expressed its protein in E. coli. DH5 alpha after inserting it in the vector and determining its nucleotide sequence. Consequentially, we attempted to prove the immunogenicity of expresion products and construct a method for detecting antibodies against GAD65 in Diabetic serum.


RESULTS
The sequence analysis showed that the amplified fragments contained 1758 bp, encoded 585 amino acid, and had been correctly inserted into pGEX-3X vector. The recombinant proteins expressed in E. Coli. DH5 alpha had immunogenicity and could be used to detect agtibodies against GAD65 in diabetic serum.


CONCLUSIONS
We have got recombinant GAD65 proteins which have immunogenicity and have used them to detect preliminarily antibodies against GAD65 in diabetic serum.
General Hospital, Tianjin Medical University.
R
R
Wu
J
J
Cai
C
C
Zhao
D000293:Adolescent; D000328:Adult; D003922:Diabetes Mellitus, Type 1 / Q000175:diagnosis*; D005968:Glutamate Decarboxylase / Q000276:immunology; D006801:Humans; D011993:Recombinant Fusion Proteins / Q000276:immunology
D011993:Recombinant Fusion Proteins; D005968:Glutamate Decarboxylase