An Efficient Ligation Method in the Making of an in vitro Virus for in vitro Protein Evolution View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2002-06

AUTHORS

Ichiro Tabuchi, Sayaka Soramoto, Miho Suzuki, Koichi Nishigaki, Naoto Nemoto, Yuzuru Husimi

ABSTRACT

The "in vitro virus" is a molecular construct to perform evolutionary protein engineering. The "virion (=viral particle)" (mRNA-peptide fusion), is made by bonding a nascent protein with its coding mRNA via puromycin in a test tube for in vitro translation. In this work, the puromycin-linker was attached to mRNA using the Y-ligation, which was a method of two single-strands ligation at the end of a double-stranded stem to make a stem-loop structure. This reaction gave a yield of about 95%. We compared the Y-ligation with two other ligation reactions and showed that the Y-ligation gave the best productivity. An efficient amplification of the in vitro virus with this "viral genome" was demonstrated. More... »

PAGES

49-54

Identifiers

URI

http://scigraph.springernature.com/pub.10.1251/bpo33

DOI

http://dx.doi.org/10.1251/bpo33

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1000044128

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/12734569


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