Introducing StuI sites improves vectors for the expression of fusion proteins with factor Xa cleavage sites.
The Arg-encoding triplet (AGG) in the recognition sequence Ile-Glu-Gly-Arg for factor Xa can be used to generate a StuI restriction site (AGGCCT) which greatly facilitates the construction of DNA fragments encoding fusion proteins. Following proteolytic cleavage with factor Xa, a protein with the desired N terminus can be obtained.
Görlach
J
J
Institute of Plant Sciences, Swiss Federal Institute of Technology, Zürich, Switzerland.
Schmid
J
J
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods*; D004274:DNA, Recombinant / Q000235:genetics*; D015252:Deoxyribonucleases, Type II Site-Specific; D004926:Escherichia coli / Q000235:genetics; D015951:Factor Xa / Q000378:metabolism*; D005822:Genetic Vectors / Q000235:genetics*; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D013379:Substrate Specificity
D004274:DNA, Recombinant; D011993:Recombinant Fusion Proteins; C028868:AGGCCT-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific; D015951:Factor Xa