Summary
As the initial step in a project to provide a more cost-effective source of the phytase enzyme, this paper reports on the use of a polyclonal antibody raised to phytase purified from an isolate of Aspergillus niger (A. ficuum) to screen an A. niger lambda gt11 expression library and the use of amino acid sequencing to identify a clone containing part of the fungal phytase gene. The described use of amino acid sequence fragments to verify the cloning of a gene has potential applications in other cloning projects.
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References
Garber RC, Yoder OC (1983) Isolation of DNA from filamentous fungi and separation into nuclear, mitochondrial, ribosomal, and plasmid components. Anal Biochem 135:416–422
Innis MA, Holland MJ, McCabe PC, Cole GE, Wittmann VP, Tal R, Watt KWK, Gelfand DH, Holland JP, Meade JH (1985) Expression, glycosylation, and secretion of an Aspergillus glucoamylase by Saccharomyces cerevisiae. Science 228:21–26
Klich MA, Mullaney EJ (1987) DNA restriction enzyme fragment polymorphism as a tool for rapid differentiation of Aspergillus flavus from Aspergillus oryzae. Exp Mycol 11:170–175
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 27:680–685
Nelson TS, Shieh TR, Wodzinski RJ, Ware JH (1971) Effect of supplemental phytase on the utilization of phytase phosphorus by chicks. J Nutr 101:1289–1294
Reddy NR, Sathe SK, Salunkhe DK (1982) Phytates in legumes and cereals. Adv Food Res 28:1–92
Shieh TR, Ware JH (1968) Survey of microorganisms for the production of extracellular phytase. Appl Microbiol 6:1348–1351
Tarr G (1986) Manual Edman sequencing systems. In: Shively J (ed) Methods of protein microcharacterization. Humana Press, Clifton, N. J., pp 155–194
Thomsen J, Bayne S (1988) Microscale alkylation with 4-vinlypyridine. J Protein Chem 7:295–296
Ullah A (1988) Production, rapid purification and catalytic characterization of extracellular phytase from Aspergillus ficuum. Prep Biochem 18:443–458
Ullah A, Cummins BJ (1988) Aspergillus ficuum extracellular pH 6.0 optimum acid phosphatase: purification, N-terminal amino acid sequence, and biochemical characterization. Prep Biochem 18:37–65
Ullah A, Gibson DM (1987) Extracellular phytase (E.C. 3.1.8.8) from Aspergillus ficuum NRRL 3135: purification and characterization. Prep Biochem 17:63–91
Young RA, Davis RW (1985) Immunoscreening lambda gt11 recombinant DNA expression libraries. In: Setlow JK, Hollaender A (eds) Genetic engineering principles and methods, vol. 7. Plenum Press, New York, pp 29–41
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Mention of a trademark or proprietary product does not constitute a guarantee or warranty by the U.S. Department of Agriculture and does not imply approval to the exclusion of other products that may also be suitable
Offprint requests to: E. Mullaney
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Mullaney, E.J., Gibson, D.M. & Ullah, A.H.J. Positive identification of a lambda gt11 clone containing a region of fungal phytase gene by immunoprobe and sequence verification. Appl Microbiol Biotechnol 35, 611–614 (1991). https://doi.org/10.1007/BF00169625
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DOI: https://doi.org/10.1007/BF00169625