A New Method for Purifying Lambda DNA From Phage Lysates

Helms, Cynthia; Graham, Madge Y.; Dutchik, James E.; Olson, Maynard V.

doi:10.1089/dna.1985.4.39

Cited by 176 publications

(68 citation statements)

References 15 publications

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“…One clone, XCIE, contained an insert that hybridized strongly to both probes. Phage DNA was prepared as described (43 19. apoE. The chromosomal location ofAPOE was determined by examining a panel of human-mouse cell hybrids with a cloned human apoE cDNA probe.…”

Section: Methodsmentioning

confidence: 99%

Regional mapping of human chromosome 19: organization of genes for plasma lipid transport (APOC1, -C2, and -E and LDLR) and the genes C3, PEPD, and GPI.

Lusis¹,

Heinzmann²,

Sparkes³

et al. 1986

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

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We report the regional mapping of human chromosome 19 genes for three apolipoproteins and a lipoprotein receptor as well as genes for three other markers. The regional mapping was made possible by the use of a reciprocal whole-arm translocation between the long arm of chromosome 19 and the short arm of chromosome 1. Examination of three separate somatic cell hybrids containing the long arm but not the short arm of chromosome 19 indicated that the genes for apolipoproteins CI, CII, and E (APOCI, APOC2, and APOE, respectively) and glucose-6-phosphate isomerase (GPI) reside on the long arm, whereas genes for the low density lipoprotein receptor (LDLR), complement component 3 (C3), and peptidase D (PEPD) reside on the short arm. When taken together with previous studies, our results suggest the following physical gene map: pter-LDLR-C3-pl3.2-PEPD-centromere-(APOE, APOCI, APOC2, GPI)-qter. In addition, we have isolated a single A phage carrying both APOCI and part ofAPOE. These genes are tandemly oriented and are separated by about 6 kilobases of genomic DNA. Since previous family studies indicate tight linkage of APOE and APOC2, the apolipoprotein genes APOCI, APOC2, and APOE form a tight complex on the long arm of chromosome 19, suggesting the possibility of coordinate regulation.

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Section: Methodsmentioning

confidence: 99%

Regional mapping of human chromosome 19: organization of genes for plasma lipid transport (APOC1, -C2, and -E and LDLR) and the genes C3, PEPD, and GPI.

Lusis¹,

Heinzmann²,

Sparkes³

et al. 1986

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

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“…Lambda DNA was prepared from plate lysates purified on DEAE cellulose columns according to the method of Helms et al (1985). DNA in amounts sufficient for both insert size determination and subcloning was obtained by collecting phage lysates from three 8.5-cm agar plates.…”

Section: Isolation Off3 Homologous Cdna Clones and Sequence Analysismentioning

confidence: 99%

The mouse neuronal cell surface protein F3: a phosphatidylinositol-anchored member of the immunoglobulin superfamily related to chicken contactin.

Gennarini

Cibelli

Rougon

et al. 1989

The Journal of Cell Biology

259

166

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Abstract. Several members of the Ig superfamily are expressed on neural cells where they participate in surface interactions between cell bodies and processes. Their Ig domains are more closely related to each other than to Ig variable and constant domains and have been grouped into the C2 set. Here, we report the cloning and characterization of another member of this group, the mouse neuronal cell surface antigen F3. The F3 eDNA sequence contains an open reading frame that could encode a 1,020-amino acid protein consisting of a signal sequence, six Ig-like domains of the C2 type, a long premembrane region containing two segments that exhibit sequence similarity to fibronectin type III repeats and a moderately hydrophobic COOH-terminal sequence. The protein does not contain a typical transmembrane segment but appears to be attached to the membrane by a phosphatidylinositol anchor. Antibodies against the F3 protein recognize a prominent 135-kD protein in mouse brain. In fetal brain cultures, they stain the neuronal cell surface and, in cultures maintained in chemically defined medium, most prominently neurites and neurite bundles. The mouse f3 gene maps to band F of chromosome 15. The gene transcripts detected in the brain by F3 cDNA probes are developmentally regulated, the highest amounts being expressed between 1 and 2 wk after birth.The F3 nucleotide and deduced amino acid sequence show striking similarity to the recently published sequence of the chicken neuronal cell surface protein contactin. However, there are important differences between the two molecules. In contrast to F3, contactin has a transmembrane and a cytoplasmic domain. Whereas contactin is insoluble in nonionic detergent and is tightly associated with the cytoskeleton, about equal amounts of F3 distribute between buffer-soluble, nonionic detergent-soluble, and detergent-insoluble fractions. Among other neural cell surface proteins, F3 most resembles the neuronal cell adhesion protein L1, with 25 % amino acid identity between their extracellular domains. Based on its structural similarity with known cell adhesion proteins of nervous tissue and with L1 in particular, we propose that F3 mediates cell surface interactions during nervous system development.

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“…All organisms synthesize a few evolutionary conserved proteins (called heat-shock proteins or HSP) in response to various harmful environmental stresses [l, 21. The most abundant and conserved HSP has a molecular mass of about 70 kDa.…”

mentioning

confidence: 99%

Molecular cloning and characterization of a constitutively expressed heat‐shock‐cognate hsc71 gene from rainbow trout

Zafarullah

Wiśniewski

Shworak

et al. 1992

European Journal of Biochemistry

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A rainbow trout major heat-shock-protein-like gene (hsp 70) and corresponding cDNA clones were isolated by hybridization to heterologous hsp70 probes. D N A sequencing revealed that this gene is structurally similar to a mammalian heat-shock-cognate hsc70 gene and consists of eight introns. Northern blot and primer extension analyses showed that the corresponding mRNA is constitutively abundant in different trout tissues and salmonid cell lines. Fragments of the isolated gene containing the -900 -+ 30 and -217 -+ 58 sequence were linked to a bacterial chloramphenicol acetyltransferase reporter gene and transiently transfected into salmonid cells. The expression pattern of these constructs supports our conclusion that the isolated genomic and cDNA clones correspond to a trout heat-shock-cognate hsc70 gene.All organisms synthesize a few evolutionary conserved proteins (called heat-shock proteins or HSP) in response to various harmful environmental stresses [l, 21. The most abundant and conserved HSP has a molecular mass of about 70 kDa. In eukaryotes, HSP70 proteins are encoded by a few heat-inducible hsp70 genes and usually their basal expression is negligible. Howcver, even without stress eukaryotic cells contain high levels of structurally related heat-shock-cognate proteins (HSC) encoded by constitutively active hsc70 genes [3, 41. The grp78 gene coding for glucose-regulated protein [5] and two mammalian testis-specific hsp70-related genes [6 -81 represent other members of that so-called hsp70 multigene family.Major information on the hsp70 gene family slructurc and regulation was accumulated from studies conducted on mammals, fruit fly and yeast. However, little is known about such genes in fish which arc important economically and are affected by such adverse environmental changes as water pollution. Iri vifro studies showed that cultured cells of chinook salmon and rainbow trout synthesize HSP70 and corresponding inRNA in response to hyperthermia, heavy metal ions and sodium arseiiite [9 -131. So far, two partial cDNA clones were obtained from arsenite-treated rainbow trout gonad (RTG-2) cells which show extensive identity with fruit fly and yeast hsp70 genes [12]. Our studies on salmonid cells revealed a high basal level of the 70-kDa band on SDS protein gels corresponding to the position of HSP70 accumulated after hyperthermia [13]. To obtain a further insight into molecular mechanisms involved in the constitutive and stress-inducible regulation of the rainbow trout (Salmo gairdneri) hsp70 C'orrrsponderirt~ to

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A New Method for Purifying Lambda DNA From Phage Lysates

Cited by 176 publications

References 15 publications

Regional mapping of human chromosome 19: organization of genes for plasma lipid transport (APOC1, -C2, and -E and LDLR) and the genes C3, PEPD, and GPI.

Regional mapping of human chromosome 19: organization of genes for plasma lipid transport (APOC1, -C2, and -E and LDLR) and the genes C3, PEPD, and GPI.

The mouse neuronal cell surface protein F3: a phosphatidylinositol-anchored member of the immunoglobulin superfamily related to chicken contactin.

Molecular cloning and characterization of a constitutively expressed heat‐shock‐cognate hsc71 gene from rainbow trout

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