Abstract
A cDNA of a mutant (K151E, R154G) of single chain urokinase-type plasminogen activator (mscu-PA) was constructed to include the natural scu-PA signal peptide sequences and transferred into the genome of Bombyx mori nuclear polyhedrosis virus (BmNPV) by transfer vectors pBE284 (derived from BmNPV) and pVL1392 (from AcNPV), respectively. Both Bombyx mori (BmN) cells and silkworm larvae were infected with the two recombinant viruses. Fibrin-plate assay showed that the re-virus from pVL1392 increased the yield of mscu-PA three times compared with the re-virus from pBE284.
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References
Bachmann F (1987). Fibrinolysis. In: Verstraete J, Vermylen HR, Arnout J, eds. Thrombosis and Haemeostasis. Leuven: Leuven University Press, pp. 227–265.
Felgner PL, Gadek TR, Holm M, Roman R, Chan HW, Wenz M, Northrop JP, Ringold GM, Danielsen M (1987) Proc. Natl. Acad. Sci. USA. 84: 7413–7417.
Gunzler WA, Steffens GJ, Otting F, Kim SMA, Frankus E, Folhe L (1982) Hoppe-Seylers Z. Physiol. Chem. 363: 1155–1165.
Iatrou K, Ito K, Witkiewicz H (1985) J. Virol. 54: 436–445.
Jarvis DL, Oker-Blom C, Summers MD (1990) J. Cell. Biochem. 42: 181–191
Liu JN, Lu L, Gao X, Zhu DX (1991) Sci. China (series B) 10: 1035–1039.
Meada S (1989) Gene transfer vectors of a baculovirus, Bombyx mori nuclear polyhedrosis virus, and their use for expression of foreign genes in insect cells. In: Mitsuhashi J, ed. Invertebrate Cell System Applications, Vol. I. Boca Raton, FL: CRC Press, pp. 167–182.
Nobuhara M, Sakamaki M, Ohnisk H, Suzuki Y (1981) J. Biochem. Jap. 90: 225–232.
O'Reilly DR, Miller LK, Luckow VA (1992) Baculovirus Expression Vectors, A Laboratory Manual. New York, NY: Freeman.
Peng GH, Ma Z, Yu RR, Xue YM, Zhu DX (1997a) Chinese Sci. Bull. 42: 972–976.
Peng GH, Ma Z, Xue YM, Chen YH, Zhu DX (1997b) Acta Biochim. Biophys. Sinica 29: 547–552.
Peng GH, Ma Z, Kuai LT, Zhu DX (1997c) Biochem. Mol. Biol. Int. 41: 887–894.
Ploug J, Kjeldgaard NO (1957) Biochim. Biophys. Acta. 24: 278–283.
Qiu P, Ding Y, Qin JC, Han KK, Zhu DX (1994) Biol. Chem. Hoppe-Seyler 375: 413–418.
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular Cloning: A Laboratory Manual. New York: Cold Spring Harbor Laboratory Press.
Steffens GJ, Gunzler WA, Otting F, Frankus E, Folhe L (1982). Hoppe-Seylers' Z. Physiol. Chem. 363: 1043–1058.
Summers MD, Smith GE (1987) A manual of methods for baculovirus vectors and insect cell culture procedures. Texas Agricultural Experiment Station Bulletin No. 1555.
Vetterlein D, Bell TE, Young PL, Robin R (1980) J. Biol. Chem. 255: 3665–3672.
White WF, Barlow GH, Mozen MM (1966) Biochemistry 5: 2160–2169.
Wun TC, Ossowski L, Reich E (1982) J. Biol. Chem. 257: 7267–7268.
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Peng, R., Yu, Z., Zang, Y. et al. High level expression of a mutant (K151E, R154G) of single chain urokinase-type plasminogen activator in silkworm. Biotechnology Letters 21, 979–985 (1999). https://doi.org/10.1023/A:1005606531948
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DOI: https://doi.org/10.1023/A:1005606531948