摘要:

A simple and fast (2–3 d) method is described for the extraction and restriction endonuclease digestion of bacterial chromosomal DNA in low melting point agarose pellets. The technique minimized the random mechanical shearing of DNA caused by conventional preparative methods and improved the resolution of electrophoretic band patterns, particularly for the smaller (<10 kb) fragments. The method was tested on 14 strains ofProvidencia stuartiiandProv. rustigiani

ISSN:0266-8254    

DOI:10.1111/j.1472-765X.1987.tb01610.x

出版商:Blackwell Publishing Ltd    

年代:1987

数据来源: WILEY

摘要:

Transmissible multiple resistance to tetracycline, ampicillin and streptomycin was found to be associated with transfer of the larger of the two plasmids (˜ 80 and ˜ 45 kbp) present in a group of tetracycline‐resistantEscherichia colistrains isolated from the rumen of sheep. A second group of rumen tetRstrains, which differed in the ability to utilize several sugars, showed non‐transmissible resistances to tetracycline and strepto

ISSN:0266-8254    

DOI:10.1111/j.1472-765X.1987.tb01611.x

出版商:Blackwell Publishing Ltd    

年代:1987

数据来源: WILEY

摘要:

We describe a rapid procedure for extracting purified chromosomal DNA fromPseudomonas putida, and some minor modifications needed for use in other organisms, including Gram‐positive strains. The technique is rapid and generally produces between 1 and 5 μg of DNA from 1 ml of liquid culture. The DNA is highly purified and can be readily cut with small quantities of restriction endo‐nucleases, cloned into plasmid vectors and used as a substrate for hybridization with labelled DNA probes. Genetical analysis and cloning of genomic DNA from environmentally important organisms likePs. putidahas become increasingly important. Bacterial chromosomal DNA is usually prepared from large volumes of liquid culture, and involves time‐consuming steps necessary to purify the DNA sufficiently for use in cloning experiments (e. g. Marmur 1961). We have developed a method for preparing small quantities of genomic DNA, which involves whole cell lysis and purification by precipitation and centrifugation. Loss of DNA occurs by shearing, but the yield of DNA is sufficient, and the quality is high. The method has been used to prepare DNA from a variety of organisms and is particularly applicable where the preparation and analysis of DNA from a large number of isolates is re

ISSN:0266-8254    

DOI:10.1111/j.1472-765X.1987.tb01612.x

出版商:Blackwell Publishing Ltd    

年代:1987

数据来源: WILEY

摘要:

A rapid, simple and sensitive technique was developed for the immunodetection and immunopurification of enterotoxins from contaminated materials. This technique, which avoids interference by protein A, involves the application of either staphylo‐coccal culture supernatant fluids or contaminated materials to an immunoabsorbent of rabbit immunoglobulins coupled to a solid matrix followed by passage through an anti‐enterotoxin antibody affinity column. The total elimination of the cross‐reactivity caused by protein A was monitored by immunoblotting with monoclonal anti‐enterotoxin ant

ISSN:0266-8254    

DOI:10.1111/j.1472-765X.1987.tb01613.x

出版商:Blackwell Publishing Ltd    

年代:1987

数据来源: WILEY

摘要:

A rapid passive haemagglutination (PHA) slide test has been developed for serotyping campylobacters on the basis of heat‐stable antigens. Chicken erythrocytes are sensitized with hot saline extracts of campylobacters and tested on a slide with staphylococci coated with antibody. Serotyping using the slide PHA system gives results which are in almost complete agreement with those obtained using a standard PHA techniqu

ISSN:0266-8254    

DOI:10.1111/j.1472-765X.1987.tb01614.x

出版商:Blackwell Publishing Ltd    

年代:1987

数据来源: WILEY