Cloning in single-stranded bacteriophage as an aid to rapid DNA sequencing

Sanger, F.; Coulson, Alan; Barrell, Bart; Smith, Andrew J.H.; Roe, Bruce A.

doi:10.1016/0022-2836(80)90196-5

Cited by 3,415 publications

(1,273 citation statements)

References 24 publications

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“…The small and large fragments were then cross-ligated to produce the chimeric constructs AB 159 -214 A (amino acid segment 159 -214 of MAO A replaced by the corresponding MAO B segment) and BA 150 -205 B. The successful construction of the point mutants and the chimerics was confirmed by dideoxy sequencing analysis (Sanger et al, 1980).…”

Section: Construction Of Ab 159 -214 a And Ba 150 -205 B Chimeric Enzmentioning

confidence: 99%

Phe²⁰⁸ and Ile¹⁹⁹ in Human Monoamine Oxidase A and B Do Not Determine Substrate and Inhibitor Specificities as in Rat

Geha

Chen

Shih

2000

Journal of Neurochemistry

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It has been reported previously that reciprocally switching Phe 208 and Ile 199 in rat monoamine oxidase (MAO) A and B, respectively, was sufficient to switch their substrate and inhibitor preferences. In this study, the same mutants were made in the human forms of MAO. When compared with MAO A, MAO A-F208I showed a sixfold decrease in the specificity constant k cat /K m for both the MAO A-and the MAO B-preferring substrates 5-hydroxytryptamine and ␤-phenylethylamine, respectively. The reciprocal point mutant MAO B-I199F had no effect on substrate affinity. To investigate if the region neighboring these two residues is responsible for conferring preferences, we have also made chimeric constructs by reciprocally switching the corresponding amino acid segments 159 -214 in 1304 -1309 (2000).Monoamine oxidase [MAO; amine:oxygen oxidoreductase (deaminating) (flavin-containing), EC 1.4.3.4] catalyzes the oxidative deamination of biogenic and xenobiotic amines and plays an important role in regulating their levels. It is a flavin-adenine dinucleotidecontaining enzyme located on the mitochondrial outer membrane (Nara et al., 1966;Greenawalt and Schnaitman, 1970;Shih et al., 1999). It exists as two isoenzymes, A and B. MAO A preferentially oxidizes serotonin [5-hydroxytryptamine (5-HT)] and is inhibited by low concentrations of clorgyline, whereas MAO B preferentially oxidizes low concentrations of ␤-phenylethylamine (PEA) and is inhibited by low concentrations of deprenyl. Common substrates include tyramine (Johnston, 1968;Knoll and Magyar, 1972) and dopamine. MAO A and B are composed of 527 and 520 amino acids, respectively, and have a 70% amino acid identity (Bach et al., 1988). They are closely linked on the X chromosome (Lan et al., 1989) and have an identical intron-exon organization, indicating that they are derived from a common ancestral gene (Grimsby et al., 1991). Knocking out the MAO A gene resulted in aggressive behavior in mice (Cases et al., 1995), whereas knocking out the MAO B gene resulted in mice resistant to the parkinsonism-inducing toxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (Grimsby et al., 1997).To determine the region(s) responsible for the distinct substrate and inhibitor preferences of the two isoenzymes, we and other groups have made point mutants and chimeric MAO constructs exchanging corresponding portions of the isoenzymes (Gottowik et al., 1993(Gottowik et al., , 1995Wu et al., 1993;Tsugeno et al., 1995;Cesura et al., 1996;Chen et al., 1996). It has been shown that amino acid segments 161-375 in human MAO A and 152-366 in human MAO B contain part of the domain responsible for determining preference (Grimsby et al., 1996). It was later discovered that reciprocally switching amino acids F208 in rat MAO A and its corresponding residue in rat MAO B, I199, was sufficient to switch the substrate and inhibitor preferences of rat MAO A and B (Tsugeno and Ito, 1997). To determine if the human MAO equivalents of these two residues, which are located within the segments in our ea...

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Section: Construction Of Ab 159 -214 a And Ba 150 -205 B Chimeric Enzmentioning

confidence: 99%

Phe²⁰⁸ and Ile¹⁹⁹ in Human Monoamine Oxidase A and B Do Not Determine Substrate and Inhibitor Specificities as in Rat

Geha

Chen

Shih

2000

Journal of Neurochemistry

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“…$1 mapping was carried out according to Bibb et al (1986) at a hybridization temperature of 45 ° C. RNA was isolated as described in Hopwood et al (1985) from cultures of S. lividans TK24 harbouring pDMLl10, grown in YEME containing 34% sucrose, 5 mM MgC12 and 0.5% glycine, for 96 h, after which time the production of DD-peptidase was maximal. Nucleotide sequencing was carried out using the M13 dideoxynucleotide (Sanger et al 1977(Sanger et al , 1980 and the Maxam and Gilbert (1980) .... …”

Section: Methodsmentioning

confidence: 99%

Transcriptional analysis of the dd-peptidase/penicillin-binding protein-encoding dac gene of Streptomyces R61: Use of the promoter and signal sequences in a secretion vector

et al. 1990

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Summary. The promoter region of the gene encoding the extracellular DD-peptidase/penicillin-binding protein of Streptomyces R61 has been identified by in vivo promoter probing and SI mapping. A secretion vector, pDML116, was constructed by inserting into the multicopy Streptomyces plasmid pIJ702, a 247 bp DNA sequence that contained the transcriptional, translational and secretory signals and the 12 amino acid N-terminal region-encoding sequence of the mature Streptomyces DD-peptidase/penicillin-binding protein. Insertion, downstream of this 247 bp segment, of the Streptomyces R61 DD-peptidase-encoding gene or the Escherichia coli R-TEM /%lactamase-encoding gene yielded plasmids pDML120 and pDML128, respectively, which allowed expression and secretion of the relevant enzymes by Streptomyces lividans. The maximal secretion levels obtained were 42 mg protein/ml for the autologous Streptomyces DD-peptidase and 0.9 mg protein/ml for the heterologous E. coli /%lactamase.

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“…Preparation of M13 mp2 DNA was carried out as in [16], except that phage were precipitated by the addition of polyethylene glycol [17]. Chain terminator sequencing reactions and analyses of the products were performed as in [8].…”

Section: Dna Sequencingmentioning

confidence: 99%

Use of a rapid DNA sequencing system to demonstrate the induction of frameshift mutations by bleomycin

1982

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The rapid DNA sequencing system based on the single-stranded bacteriophage M 13 and the chain-terminator method has been used to look directly for mutational alterations. A small DNA fragment that primes DNA synthesis through the N-terminal 200 base pairs of the fl-galactosidase gene was prepared, and used to detect changes in base sequence among phages that give white plaques after treatment of the host cells with bleomycin. Bleomycin treatment of E. coli in which M 13 mp2 was growing gave an increase in white plaque frequency. DNA sequence analysis of phage from 7 independent mutant plaques showed them all to have a frameshift mutation.

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Cloning in single-stranded bacteriophage as an aid to rapid DNA sequencing

Cited by 3,415 publications

References 24 publications

Phe²⁰⁸ and Ile¹⁹⁹ in Human Monoamine Oxidase A and B Do Not Determine Substrate and Inhibitor Specificities as in Rat

Phe²⁰⁸ and Ile¹⁹⁹ in Human Monoamine Oxidase A and B Do Not Determine Substrate and Inhibitor Specificities as in Rat

Transcriptional analysis of the dd-peptidase/penicillin-binding protein-encoding dac gene of Streptomyces R61: Use of the promoter and signal sequences in a secretion vector

Use of a rapid DNA sequencing system to demonstrate the induction of frameshift mutations by bleomycin

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Cloning in single-stranded bacteriophage as an aid to rapid DNA sequencing

Cited by 3,415 publications

References 24 publications

Phe208 and Ile199 in Human Monoamine Oxidase A and B Do Not Determine Substrate and Inhibitor Specificities as in Rat

Phe208 and Ile199 in Human Monoamine Oxidase A and B Do Not Determine Substrate and Inhibitor Specificities as in Rat

Transcriptional analysis of the dd-peptidase/penicillin-binding protein-encoding dac gene of Streptomyces R61: Use of the promoter and signal sequences in a secretion vector

Use of a rapid DNA sequencing system to demonstrate the induction of frameshift mutations by bleomycin

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Phe²⁰⁸ and Ile¹⁹⁹ in Human Monoamine Oxidase A and B Do Not Determine Substrate and Inhibitor Specificities as in Rat

Phe²⁰⁸ and Ile¹⁹⁹ in Human Monoamine Oxidase A and B Do Not Determine Substrate and Inhibitor Specificities as in Rat