Mouse immunoglobulin genes: a bacterial plasmid containing the entire coding sequence for a pre-γ 2a heavy chain

Abstract

A DNA sequence complementary to the entire coding part of a mouse gamma 2a immunoglobulin heavy chain mRNA isolated from a myeloma producing a levan binding protein (UPC 10), has been cloned in the PstI site of pBR 322. Transformants containing sequences complementary to purified gamma 2a heavy chain mRNA were selected. One transformant, pG2a-10-21, containing a 1750 nucleotide insert, has been characterized by hybrid-arrested translation and purification of gamma 2a heavy chain mRNA on DNA-DBM cellulose filters. Restriction enzyme analysis and partial sequencing demonstrate that the pG2a-10-21 contains the complete structural sequence for the gamma 2a heavy chain and predicts the sequence of a 18 amino acid hydrophobic amino terminal extra piece segment.

Cloning of an intracellular receptor for protein kinase C: a homolog of the beta subunit of G proteins.

“…The nitrocellulose pieces (containing PKC/RACK1 complexes) were incubated with sample buffer and subjected to SDS/PAGE to elute the bound proteins. Western [19][20][21][22][23][24][25][26][27][28][29][30][31][32][33][34][35][36] were synthesized at the Biomolecular Resource Center, University of California, San Francisco. The purity and structural integrity of the peptides were confirmed by HPLC, amino acid analysis, and fast-atom bombardment mass spectroscopy.…”

Different isoforms and stock‐specific variants of the cell adhesion molecule C‐CAM (cell‐CAM 105) in rat liver

“…In brief, total RNA was extracted from rat liver and lung by a phenolkhloroform method [26,271 and passed through an oligo(dT)-cellulose column (Pharmacia LKB) according to [28] for the purification of poly(A)-rich RNA. The oligonucleotide probes were radiolabeled by [a-"PIdATP (6000 Ci/mmol; New England Nuclear) using a DNA tailing kit (Boehringer Mannheim).…”

Characterization of rat brain NCAM mRNA using DNA oligonucleotide probes

“…Total RNA was extracted from rat brain and rat liver by a phenolchloroform method [16,17] and then passed through an oligo(dT)-cellulose column (Pharmacia) according to [18].…”