Source:http://linkedlifedata.com/resource/pubmed/id/17544364
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
3
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pubmed:dateCreated |
2007-6-18
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pubmed:abstractText |
Bombyxin (bx) and prophenoloxidase-activating enzyme (ppae) signal peptides from Bombyx mori, their modified signal peptides, and synthetic signal peptides were investigated for the secretion of GFP(uv)-beta1,3-N-acetylglucosaminyltransferase 2 (GGT2) fusion protein in B. mori Bm5 cells and silkworm larvae using cysteine protease deficient B. mori multiple nucleopolyhedrovirus (BmMNPV-CP(-)) and its bacmid. The secretion efficiencies of all signal peptides were 15-30% in Bm5 cells and 24-30% in silkworm larvae, while that of the +16 signal peptide was 0% in Bm5 cells and 1% in silkworm larvae. The fusion protein that contained the +16 signal peptide was expressed specifically in the endoplasmic reticulum (ER) and in the fractions of cell precipitations. Ninety-four percent of total intracellular beta1,3-N-acetylglucosaminyltransferase (beta3GnT) activity was detected in cell precipitations following the 600, 8000, and 114,000g centrifugations. In the case of the +38 signal peptide, 60% of total intracellular activity was detected in the supernatant from the 114,000g spin, and only 1% was found in the precipitate. Our results suggest that the +16 signal peptide might be situated in the transmembrane region and not cleaved by signal peptidase in silkworm or B. mori cells. Therefore, the fusion protein connected to the +16 signal peptide stayed in the fat body of silkworm larvae with biological function, and was not secreted extracellularly.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Green Fluorescent Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Isoenzymes,
http://linkedlifedata.com/resource/pubmed/chemical/N-Acetylglucosaminyltransferases,
http://linkedlifedata.com/resource/pubmed/chemical/Protein Sorting Signals,
http://linkedlifedata.com/resource/pubmed/chemical/Recombinant Fusion Proteins
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pubmed:status |
MEDLINE
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pubmed:month |
Aug
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pubmed:issn |
0006-291X
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:day |
3
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pubmed:volume |
359
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
543-8
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pubmed:meshHeading |
pubmed-meshheading:17544364-Amino Acid Sequence,
pubmed-meshheading:17544364-Animals,
pubmed-meshheading:17544364-Bombyx,
pubmed-meshheading:17544364-Cell Line,
pubmed-meshheading:17544364-Fat Body,
pubmed-meshheading:17544364-Gene Expression,
pubmed-meshheading:17544364-Green Fluorescent Proteins,
pubmed-meshheading:17544364-Isoenzymes,
pubmed-meshheading:17544364-Larva,
pubmed-meshheading:17544364-Molecular Sequence Data,
pubmed-meshheading:17544364-Mutagenesis,
pubmed-meshheading:17544364-N-Acetylglucosaminyltransferases,
pubmed-meshheading:17544364-Protein Sorting Signals,
pubmed-meshheading:17544364-Recombinant Fusion Proteins
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pubmed:year |
2007
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pubmed:articleTitle |
Specific expression of GFPuv-beta1,3-N-acetylglucosaminyltransferase 2 fusion protein in fat body of Bombyx mori silkworm larvae using signal peptide.
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pubmed:affiliation |
Laboratory of Biotechnology, Faculty of Agriculture, Shizuoka University, Shizuoka 422-8529, Japan.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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