A novel method for cloning of coding sequences of highly toxic proteins

• Published in: Biochimica et biophysica acta. General subjects Vol. 1863; no. 3; pp. 521 - 527
• Main Authors: Krela, Rafal, Poreba, Elzbieta, Weglewska, Martyna, Skrzypczak, Tomasz, Lesniewicz, Krzysztof
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.03.2019
• Subjects: Acanthamoeba castellanii; bacteria; bacterial growth; cell proliferation; complementary DNA; deoxyribonucleases; Frameshift introduction; gene overexpression; messenger RNA; molecular cloning; Nuclease genes; nucleotide sequences; Protein overexpression; recombinant proteins; Toxic protein; toxicity; transgenes; translation (genetics); Nuclease genes; Toxic protein; Acanthamoeba castellanii; Protein overexpression; Frameshift introduction

During standard gene cloning, the recombinant protein appearing in bacteria as the result of expression leakage very often inhibits cell proliferation leading to blocking of the cloning procedure. Although different approaches can reduce transgene basal expression, the recombinant proteins, which ev...