The construction of cosmid libraries which can be used to transform eukaryotic cells

Grosveld, Frank; Lund, Torben; Murray, Edward J.; Mellor, Andrew L.; Dahl, Hans Henrik M.; Flavell, Richard A.

doi:10.1093/nar/10.21.6715

Cited by 316 publications

(141 citation statements)

References 31 publications

Supporting

Mentioning

136

Contrasting

Unclassified

Order By: Relevance

“…Two factors do appear to be of potential relevance. Our vector containing the globin -y gene contained an intact SV40 promoter enhancer element, whereas the vector studied by Wright et al (1983) contained the herpes simplex thymidine kinase gene that lacks an identifiable enhancer (Grosveld et al, 1982 (Anagnou et al, 1985b) is consistent with this lack of evolutionary conservation. Our linker-scanning mutants did not encompass both 'CCAAT' elements although deletion of the single 'CCAAT' elements from other globin gene promoters consistently reduces function several fold (Mellon et al, 1981;Dierks et al, 1983;Charnay et al, 1985).…”

Section: Discussionmentioning

confidence: 61%

Promoter sequences required for function of the human gamma globin gene in erythroid cells.

Anagnou¹,

Karlsson²,

Moulton³

et al. 1986

The EMBO Journal

View full text Add to dashboard Cite

The human 'y-and 3-globin genes are expressed during the fetal and adult developmental periods, respectively. Differences in the sequences of their promoters may be relevant to their developmental regulation. The y globin gene promoter was found to be stronger than that of the , gene. In HeLa cells co-transfected with plasmids containing the two genes, transcripts arising from the -y promoter accumulated to a level 3-fold higher than those initiated from the f-promoter. We have recently shown that deletion of the most distal of the conserved elements of the promoter (CACCC) reduces function to 25% of the wild-type, whereas the removal of the proximal of the two duplicated (CCAAT) elements increases promoter function 2-to 4-fold in HeLa cells during transient gene expression. Both the wild-type promoter and the truncation and linker-scanning mutants from which one of the two duplicated 'CCAAT' elements had been removed, exhibited regulated expression when stably integrated into chromosomes of mouse erythroleukemia (MEL) cells. Thus, duplication of the 'CCAAT' element, the feature by which the 'y promoter differs most strikingly from the (3, is not essential for function in erythroid cells.

show abstract

Section: Discussionmentioning

confidence: 61%

Promoter sequences required for function of the human gamma globin gene in erythroid cells.

Anagnou¹,

Karlsson²,

Moulton³

et al. 1986

The EMBO Journal

View full text Add to dashboard Cite

show abstract

“…In brief, DNA from the consanguineous homozygous cell line HHK was partially digested with Sau3A1 and fractionated on sucrose density gradients. DNA fractions of 35-40 kilobases (kb) were collected and ligated with arms of cosmid pOPF (12), which had been treated with phosphatase. The ligation mix was packaged in vitro (13) and introduced into Escherichia coli ED8767.…”

Section: Methodsmentioning

confidence: 99%

“…A cosmid library of digested DNA from the homozygous B-cell line HHK was constructed by using the cosmid vector pOPF (12). The library was screened by colony filter hybridization with a probe corresponding to the first domain of an HLA-DR 13-chain gene probe cloned in bacteriophage X (unpublished data).…”

Section: Mapping Of Cosmid Clones Containing the Sb /-Chainmentioning

confidence: 99%

Molecular organization of the HLA-SB region of the human major histocompatibility complex and evidence for two SB beta-chain genes.

Gorski¹,

Rollini²,

Long³

et al. 1984

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

The class II products of the major histocompatibility complex, also called Ia antigens, are composed of two polypeptide chains, the a and P chains, both encoded within the major histocompatibility complex. In man, the class II antigens can be divided into three biochemically distinct grodps called HLA-DR, HLA-DC, and HLA-SB. Our isolation of cDNA clones for the polymorphic fi chain of HLA-DR and HLA-DC has allowed us to study the organization of the class II genes. Here we identify the HLA-SB (-chain gene in recombinant clones from a cosmid library generated from a consanguineous homozygous B-cell line. The SB 3-chain gene is linked to the SB a-chain gene and the two genes are in opposite orientation. A second SB (&chain gene, corresponding to a new SB 3II locus, has also been identified and cloned. The SB 3chain genes show much less allelic restriction site polymorphism than the genes for the ( chains of HLA-DR or HLA-DC. MATERIALS AND METHODSConstruction of the Cosmid Library. Details of the cosmid library will be published elsewhere. In brief, DNA from the consanguineous homozygous cell line HHK was partially digested with Sau3A1 and fractionated on sucrose density gradients. DNA fractions of 35-40 kilobases (kb) were collected and ligated with arms of cosmid pOPF (12), which had been treated with phosphatase. The ligation mix was packaged in vitro (13) and introduced into Escherichia coli ED8767. The packaged cosmids were spread, replicated (14), and screened by colony filter hybridization (15). A DNA fragment from a DR p-chain genomic clone (unpublished data) labeled by nick-translation was used for these hybridizations. Colonies that were positive after three rounds of screening were grown and cosmid DNA was prepared.Identification of a Possible SB f3 cDNA Clone. The cloning of class II p-chain cDNA clones from a DR4,w6 cell line has been described (9 (Fig. 1) 3934The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.

show abstract

“…29,30 Genomic subfragments of Alu-positive phages or cosmids were subcloned into digested pT7T3 plasmid DNA (Amersham Pharmacia Biotech, Freiburg, Germany).…”

Section: Genomic Cloningmentioning

confidence: 99%

Activation ofgef-h1, a guanine nucleotide exchange factor for RhoA, by DNA transfection

et al. 2004

View full text Add to dashboard Cite

Several oncogenes isolated by the NIH/3T3 transformation assay, i.e., dbl, dbs, lbc, lfc, lsc, net, ost and tim, contain a Dbl homology (DH) and a pleckstrin-homology (PH) domain and act as GEFs (guanine nucleotide exchange factors) for Rho-like GTPases. In a search for genes with oncogenic potential in DNA from the monocytic leukaemia cell line U937, we identified an amino-terminal truncated form of gef-h1, a gene encoding a GEF for RhoA. These data support the idea that a systematic search for mutations and/or deletions of GEFs in human cancer is promising.

show abstract

The construction of cosmid libraries which can be used to transform eukaryotic cells

Cited by 316 publications

References 31 publications

Promoter sequences required for function of the human gamma globin gene in erythroid cells.

Promoter sequences required for function of the human gamma globin gene in erythroid cells.

Molecular organization of the HLA-SB region of the human major histocompatibility complex and evidence for two SB beta-chain genes.

Activation ofgef-h1, a guanine nucleotide exchange factor for RhoA, by DNA transfection

Contact Info

Product

Resources

About