Functional expression on bacteriophage of the mustard trypsin inhibitor MTI-2.
The mustard trypsin inhibitor MTI-2 is a potential tool in the study of interactions between pest insects and plants. It can be applied to study the adaptations of digestive proteases in pest insects. Phage display allows a rapid and exhaustive system for the selection of heterologous protein variants with novel specificities. Here we describe a bacteriophage expression system which permits functional expression of MTI-2 variants. Active and inactive mutants of MTI-2 are constructed and displayed on phage. These are used to demonstrate that an active variant can be selected from a background of 10,000 inactive mutants in four rounds of selection and amplification.
Volpicella
M
M
Dipartimento di Biochimica e Biologia Molecolare, Universita' di Bari, Via Amendola 165/A, 70126 Bari, Italy. m.volpicella@biologia.uniba.it
Ceci
L R
LR
Gallerani
R
R
Jongsma
M A
MA
Beekwilder
J
J
D000818:Animals; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D014644:Genetic Variation; D009149:Mustard Plant / Q000235:genetics*; D019151:Peptide Library; D010843:Pichia / Q000235:genetics; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D010946:Plants, Medicinal; D015202:Protein Engineering; D014361:Trypsin Inhibitors / Q000235:genetics* / Q000378:metabolism
D017931:DNA Primers; C405940:MTI2 protein, Sinapis alba; D019151:Peptide Library; D010940:Plant Proteins; D014361:Trypsin Inhibitors