In vitro reconstitution of 30S ribosomal subunits using complete set of recombinant proteins.
This system allows convenient purification of large quantities of all of the small subunit ribosomal proteins by overexpression from cloned genes. This not only allows large-scale reconstitution of 30S subunits from individual proteins, but also facilitates protein purification greatly. These proteins can be reconstituted into functional 30S subunits using an ordered assembly protocol based on the in vitro 30S assembly map. Reconstitution of 30S subunits using this system enables mutant or modified proteins, such as Fe(II)-BABE-derivatized proteins, to be incorporated into subunits for studying ribosome structure and function.
Culver
G M
GM
Department of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames 50011, USA.
Noller
H F
HF
D002853:Chromatography, Liquid; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics; D005821:Genetic Techniques; D010649:Phenylalanine / Q000737:chemistry; D010957:Plasmids / Q000378:metabolism; D012343:RNA, Transfer / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D012269:Ribosomal Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism
D011994:Recombinant Proteins; D012269:Ribosomal Proteins; C040469:ribosomal protein S30; D010649:Phenylalanine; D012343:RNA, Transfer