Protein arrays from cDNA expression libraries.
This chapter describes the production of a cDNA expression library from human fetal brain, the construction of a high-density protein array from such a library, and two applications to screen the array for binding proteins. After producing the library and decollating the expression clones, one can pick thousands of expression clones with a laboratory robot and can deposit them into microtiter plates in an ordered manner. Such ordered clone libraries are the starting material for the construction of a high-density protein array. This array is constructed by spotting the expression clones onto a protein-binding membrane. Following cell growth and induction of protein expression on the membrane, the cell spots are lysed and their recombinant protein immobilized on the membrane. The so-constructed array carries thousands of proteins without the need to clone, express, and spot individual proteins. Such arrays allow one to screen for numerous protein functions in a high-throughput manner.
Weiner
Hendrik
H
Department of Vertebrate Genomics, Max Planck Institute of Molecular Genetics, Berlin, Germany.
Faupel
Thomas
T
Büssow
Konrad
K
D000595:Amino Acid Sequence; D001483:Base Sequence; D001923:Brain Chemistry; D003001:Cloning, Molecular; D005333:Fetus / Q000033:anatomy & histology; D015723:Gene Library; D006801:Humans; D008969:Molecular Sequence Data; D040081:Protein Array Analysis; D011485:Protein Binding; D011994:Recombinant Proteins / Q000737:chemistry* / Q000378:metabolism
D011994:Recombinant Proteins