In situ synthesis of protein arrays.
In situ or on-chip protein array methods use cell free expression systems to produce proteins directly onto an immobilising surface from co-distributed or pre-arrayed DNA or RNA, enabling protein arrays to be created on demand. These methods address three issues in protein array technology: (i) efficient protein expression and availability, (ii) functional protein immobilisation and purification in a single step and (iii) protein on-chip stability over time. By simultaneously expressing and immobilising many proteins in parallel on the chip surface, the laborious and often costly processes of DNA cloning, expression and separate protein purification are avoided. Recently employed methods reviewed are PISA (protein in situ array) and NAPPA (nucleic acid programmable protein array) from DNA and puromycin-mediated immobilisation from mRNA.
He
Mingyue
M
Technology Research Group, The Babraham Institute, Cambridge CB22 3AT, UK. mingyue.he@bbsrc.ac.uk
Stoevesandt
Oda
O
Taussig
Michael J
MJ
D001709:Biotechnology; D002474:Cell-Free System; D004247:DNA / Q000235:genetics; D016133:Polymerase Chain Reaction; D040081:Protein Array Analysis / Q000379:methods*; D014176:Protein Biosynthesis; D011691:Puromycin; D012333:RNA, Messenger / Q000235:genetics
D012333:RNA, Messenger; D011691:Puromycin; D004247:DNA