Enhanced production of secretory beta1,3-N-acetylglucosaminyltransferase 2 fusion protein into hemolymph of Bombyx mori larvae using recombinant BmNPV bacmid integrated signal sequence.
The enhanced secretion of beta1,3-N-acetylglucosaminyltransferase 2 (beta3GnT2) fusion protein into the hemolymph of Bombyx mori larvae was studied using a recombinant B. mori nucleopolyhedrovirus (BmNPV) bacmid integrating seven signal sequences. When the BmNPV bacmid encoding the signal sequences from the silkworm B. mori bombyxin (bx) and B. mori prophenoloxidase-activating enzyme (ppae) was injected into silkworm larvae, 56.1 and 51.5mU/ml beta3GnT, respectively, were secreted into the hemolymph of silkworm larvae. For bx, 97.3% of the total beta3GnT activity was secreted into hemolymph, and only 1.1% remained in the intestines of silkworm larvae. For ppae, 90.8% of the total beta3GnT activity was secreted to the hemolymph, but 7.8% remained in the intestines of silkworm larvae. Using the BmNPV bacmid encoding bx, the amount of secreted beta3GnT was 91mug per larva, which was 2.5% of the total amount of protein in the hemolymph.
Park
Enoch Y
EY
Laboratory of Biotechnology, Department of Applied Biological Chemistry, Shizuoka University, 836 Ohya Suruga-Ku, Shizuoka, Japan. yspark@agr.shizuoka.ac.jp
Kageshima
Ayano
A
Kwon
Mi-Sun
MS
Kato
Tatsuya
T
D000818:Animals; D001483:Base Sequence; D012831:Bombyx / Q000201:enzymology* / Q000254:growth & development; D002460:Cell Line; D017931:DNA Primers; D005964:Glucosyltransferases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D006458:Hemolymph / Q000201:enzymology*; D007814:Larva / Q000201:enzymology; D008969:Molecular Sequence Data; D017351:N-Acetylglucosaminyltransferases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D014162:Transfection
D017931:DNA Primers; D011993:Recombinant Fusion Proteins; D005964:Glucosyltransferases; D017351:N-Acetylglucosaminyltransferases