Improved differential screening approach to analyse transcriptional variations in organized cDNA libraries.

A cDNA library was generated from rat brain tissues and organized into 1536-well plates, using a fluorescence activated cell sorter (FACS), acting as a single cell deposition system. The organized library containing 10,000 clones, with 60% full-length cDNA inserts, allowed the generation of multiple identical membrane replicas. Each replica was hybridized with a complex probe obtained from a particular brain tissue or a given cultured cell.

Interleukin-13 is a new human lymphokine regulating inflammatory and immune responses.

The discovery of new cytokines normally relies on a prior knowledge of at least one of their biological effects, which is used as a criterion either for the purification of the protein or for the isolation of the complementary DNA by expression cloning. However, the redundancy of cytokine activities complicates the discovery of novel cytokines in this way, and the pleiotropic nature of many cytokines means that the principal activities of a new cytokine may bear little relation to that used for its isolation. We have adopted an alternative approach which relies on differential screening of an organized subtracted cDNA library from activated peripheral blood mononuclear cells, using the inducibility of lymphokine messenger RNAs by anti-CD28 as a primary screening criterion.

Molecular cloning of the MCP-3 chemokine gene and regulation of its expression.

We have isolated a cDNA (NC28) transcribed from a mRNA which is transiently induced in U937 promonocytic cells by PMA and super-induced by cycloheximide. NC28 cDNA encodes a new member of the chemokine family, MCP-3, recently purified from MG-63 osteosarcoma cells by Van Damme et al. [1].

High molecular mass forms of basic fibroblast growth factor are initiated by alternative CUG codons.

A 6.75-kilobase human hepatoma-derived basic fibroblast growth factor (bFGF) cDNA was cloned and sequenced. An amino-terminal sequence generated from a purified hepatoma bFGF was found to correspond to the nucleotide sequence and to begin 8 amino acids upstream from the putative methionine start codon thought to initiate a 154-amino acid bFGF translation product.

Abundant excretion of human growth hormone by recombinant-plasmid-transformed monkey kidney cells.

A recombinant plasmid was constructed permitting the efficient synthesis of human growth hormone (hGH) in monkey cells. The plasmid contains the cDNA sequence of the hGH precursor and controlled by the SV40 early promoter, an intron, and the poly(A)-addition site of the mouse alpha-globin gene. To permit selection of transformed cells, a selectable marker (xanthine-guanine phosphoribosyl transferase; XGPRT) was also introduced into this plasmid.