Transformation of E. coli using homopolymer-linked plasmid chimeras

Peacock, Susan; Mciver, Carolyn M; Monahan, John J.

doi:10.1016/0005-2787(81)90014-9

Cited by 100 publications

(28 citation statements)

References 12 publications

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“…Prior studies (18,27,30,31,33,45,46) have demonstrated that specific domains within the 3' UT regions of the various isoactins are isotype specific. To analyze these regions of pRVaA-19 and pREyA-11, we generated two subclones designated pRVaA-3' UT-DP and pRE-yA-3' UT-XP, which contain the 3' UT regions of the rat a-vascular and -y-enteric isoactins, respectively.…”

Section: Resultsmentioning

confidence: 99%

The development expression of the rat alpha-vascular and gamma-enteric smooth muscle isoactins: isolation and characterization of a rat gamma-enteric actin cDNA.

McHugh¹,

Lessard²

1988

Mol. Cell. Biol.

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We have isolated and characterized two cDNA clones from whole rat stomach, pRVaA-19 and pRE-yA-ll, which are specific for the a-vascular and -y-enteric smooth muscle isoactins, respectively. The rat y-enteric smooth muscle actin contains a single amino acid substitution of a proline for a glutamine at position 359 of the mature peptide when compared with the chicken gizzard y-actin sequence (J. Vandekerckhove and K. Weber, FEBS Lett. 102:219, 1979). Sequence comparisons of the 5' and 3' untranslated (UT) regions of the two smooth muscle actin cDNAs demonstrate that these regions contain no apparent sequence similarities. Additional comparisons of the 5' UT regions of the two smooth muscle actin cDNAs to all other known actin sequences reveal no apparent sequence similarities for the rat -y-enteric isoactin within the 15 base pairs of sequence currently available, while the rat a-vascular isoactin contains two separate sequences which are similar to sequences within the 5' UT regions of the human and chicken a-vascular actin genes. A similar comparison of the 3' UT regions of the two smooth muscle actins demonstrates that the a-vascular isoactins do not contain the high degree of cross-species sequence conservation observed for the other isoactins and that the y-enteric isoactin contains an inverted sequence of 52 nucleotides which is similar to a sequence found within the 3' UT regions of the human, chicken, and rat 0-cytoplasmic isoactins. These observations complicate the apparent cross-species conservation of isotype specificity of these domains previously observed for the other actin isoforms. Northern blot analysis of day 15 rat embryos and newborn, day 19 postbirth, and adult rats demonstrates that the day 15 rat embryo displays low to undetectable levels of smooth muscle isoactin mRNA expression. By birth, the stomach and small intestine show dramatic increases in a-vascular and -y-enteric actin expression. These initially high levels of expression decrease through day 19 to adulthood. In the adult rat, the uterus and aorta differ in their content of smooth muscle isoactin mRNA. These results demonstrate that the y-enteric and a-vascular isoactin mRNAs are coexpressed to various degrees in tissues which contain smooth muscle.

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Section: Resultsmentioning

confidence: 99%

The development expression of the rat alpha-vascular and gamma-enteric smooth muscle isoactins: isolation and characterization of a rat gamma-enteric actin cDNA.

McHugh¹,

Lessard²

1988

Mol. Cell. Biol.

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“…In the course of our studies, ND ANTIGENS MAY SPECIFY TRANSCRIPTIONAL DOMAINS 6171 produced a fusion protein with a predicted Mr of 62,000. The TrpE-ND protein was overexpressed in Escherichia coli RR1 (3,35) as previously described (45,46). The fusion protein was excised from a preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gel and electroeluted.…”

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confidence: 99%

Nuclear dot antigens may specify transcriptional domains in the nucleus.

Xie¹,

Lambie²,

Snyder³

1993

Mol. Cell. Biol.

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A bank of 892 human autoimmune serum samples was screened by indirect immunofluorescence on human tissue culture A wide variety of cellular processes occur within the nucleus, including DNA replication, RNA transcription and processing, and ribosome biogenesis (18,33). Some of these activities are carried out in distinct regions of the nucleus; ribosome biogenesis occurs within the nucleolus (13), splicing complexes are organized into splicing domains (14, 17), and DNA replication occurs at discrete sites (15). Other less well-characterized regions of the nucleus have also been described, including nuclear bodies, which appear as granular fibrils defining the space between perichromatin and perinuclear chromatin (7, 52), and polymorphic interphase karyosomal associations, which appear as nuclear domains that are very heterogeneous in size and number from one cell to the next (42). To what extent other morphological domains exist in the nucleus and whether they carry out distinct cellular functions are not known.One method to determine whether specific subcellular domains exist within the mammalian nucleus is to screen sera from autoimmune disease patients for antibodies that recognize novel staining patterns. Patients with specific autoimmune diseases often contain antibodies directed to nuclear components (10,27,51). These include antibodies directed to kinetochores (5, 12, 20, 32), nuclear lamins (24, 37), topoisomerase I (29), proliferating-cell nuclear antigen (4,28,36), and splicing components (26,34,38). One unusual type of autoantibody recognizes speckled dots or nuclear dots (NDs) (2, 6, 39), which are visible as approximately six brightly staining dots in the nucleus.In this report, we describe autoantibodies directed to a 72,000-Mr ND antigen. The ND domain is distinct from known nuclear domains such as splicing centers or kinetochores. Cloning and molecular characterization of a major ND antigen are also reported. In the course of our studies, * Corresponding author. reports from two other laboratories have partially described ND antigens (2, 50). Our results indicate that the 72-kDa ND antigen is associated with nuclear DNA and can serve as a transcriptional activator in Saccharomyces cerevisiae and mammalian cells. MATERIALS AND METHODSStrains and sera. Human autoimmune sera were provided by J. Hardin and J. Craft of the Yale University School of Medicine and screened as described by Yang et al. (56

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“…Terminal deoxynucleotidyl transferase was from Pharmacia. Annealing of cDNA and vector was done at a vector concentration of 0.4 p.g/ml and a vector/cDNA ratio (wt/wt) of 40:1 (15,31).…”

mentioning

confidence: 99%

“…Recombinant DNA was used to transform CaC12-competent W. coli HB101 cells, and transformants were selected by plating cells on L-broth agar plates containing ampicillin at 0.1,ug/ml (31).…”

mentioning

confidence: 99%

Molecular cloning of gene sequences regulated during squamous differentiation of tracheal epithelial cells and controlled by retinoic acid.

1987

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A cDNA library was constructed from polyadenylated RNA present in squamous differentiated rabbit tracheal epithelial cells. Screening of the cDNA library was aimed at identifying RNAs that were abundant in squamous cells and expressed at low levels in undifferentiated cells. Two different recombinants were obtained containing inserts, 0.86 and 0.77 kilobases (kb) in size, that hybridized to mRNAs 1.0 and 1.25 kb in length. These RNAs were present at approximately 50-fold higher levels in squamous cells than in proliferative or confluent retinoic acid-treated cells. The increase in the levels of the 1.0-and 1.25-kb RNAs correlated closely with the onset of squamous difrerentiation and was not related to induction of terminal cell division. Treatment of rabbit tracheal epithelial cells with transforming growth factor IS, which induces squamous differentiation in these cells, also resulted in elevated levels of the 1.0-and 1.25-kb RNAs. The increased levels of these RNAs in squamous cells appeared to a large extent to be regulated at a posttranscriptional level. Retinoic acid not only inhibited the increase in the levels of the 1.0-and 1.25-kb RNAs but also reversed the expression of these RNAs in squiamous ceUs. These results suggest that retinoic acid affects, directly or indirectly, molecular events that induce alterations in the posttranscriptional processing of the transcripts corresponding to the 1.0-and 1.25-kb RNAs.

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Transformation of E. coli using homopolymer-linked plasmid chimeras

Cited by 100 publications

References 12 publications

The development expression of the rat alpha-vascular and gamma-enteric smooth muscle isoactins: isolation and characterization of a rat gamma-enteric actin cDNA.

The development expression of the rat alpha-vascular and gamma-enteric smooth muscle isoactins: isolation and characterization of a rat gamma-enteric actin cDNA.

Nuclear dot antigens may specify transcriptional domains in the nucleus.

Molecular cloning of gene sequences regulated during squamous differentiation of tracheal epithelial cells and controlled by retinoic acid.

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