Expression studies of CLN3 protein.
Expression of the gene for Batten disease (CLN3) was studied in Escherichia coli and in a cell-free rabbit reticulocyte expression systems. A full-length recombinant fusion CLN3 protein was not produced in the bacterial systems used. However, both N-terminal fragment encompassing 246 amino acids and short C-terminal fragment containing 428-438 amino acids of the CLN3 protein were successfully overexpressed in bacteria. Further studies showed that the C-terminal sequence of the CLN3 protein corresponding to the 356-438 amino acid residues was responsible for inhibition of protein synthesis in bacteria. The full-length CLN3 gene product was readily synthesized in vitro in the cell-free rabbit reticulocyte expression system. The product obtained, corresponding to core CLN3 protein, showed an approximate molecular weight of 43 kDa. Immunoprecipitation of this product with pAb to 4-19 amino acids of the CLN3 protein allows us to suggest that CLN3 protein translation starts at Met-1.
Kaczmarski
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W
Department of Pathological Neurobiology, NYS Institute for Basic Research in Developmental Disabilities, USA.
Kida
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E
Lach
A
A
Rubenstein
R
R
Zhong
N
N
Wisniewski
K E
KE
D000293:Adolescent; D000818:Animals; D002474:Cell-Free System; D002648:Child; D002675:Child, Preschool; D016213:Cyclins; D018076:DNA, Complementary / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression / Q000502:physiology; D006801:Humans; D007223:Infant; D008562:Membrane Glycoproteins; D018832:Molecular Chaperones; D009472:Neuronal Ceroid-Lipofuscinoses / Q000175:diagnosis / Q000235:genetics*; D010446:Peptide Fragments / Q000235:genetics; D014176:Protein Biosynthesis / Q000235:genetics; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000235:genetics; D012156:Reticulocytes; D029701:Saccharomyces cerevisiae Proteins
C079895:CLN3 protein, S cerevisiae; C105199:CLN3 protein, human; D016213:Cyclins; D018076:DNA, Complementary; D008562:Membrane Glycoproteins; D018832:Molecular Chaperones; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins