![]() ![]() ![]() The cookie is set by the GDPR Cookie Consent plugin and is used to store whether or not user has consented to the use of cookies. The cookie is used to store the user consent for the cookies in the category "Performance". ![]() This cookie is set by GDPR Cookie Consent plugin. The cookies is used to store the user consent for the cookies in the category "Necessary". The cookie is used to store the user consent for the cookies in the category "Other. Providing professional IT services and solutions to the Dallas-Fort Worth area including Network Data Cabling, Network & Voice Consulting, and Surveillance & Security Solutions. This message can then be triggered based on location or date and time. This cookie is set by GDPR Cookie Consent plugin. With Ntouch, you can create an initial message to say hello to all of your friends with a new type of group chat that sends messages to each individual person in the group. The cookie is set by GDPR cookie consent to record the user consent for the cookies in the category "Functional". The cookie is used to store the user consent for the cookies in the category "Analytics". These cookies ensure basic functionalities and security features of the website, anonymously. Quantities of BtgA in reaction mixtures: lanes 1 to 6, 100 ng, 300 ng, 500 ng, 1 μg, 2 μg, and 3 μg, respectively lane C, none.Necessary cookies are absolutely essential for the website to function properly. (B) Cleavage pattern of the upper strand of oriT. The inset shows the DNase I digestion pattern up to the top of the gel, indicating that no regions of protection were seen, and also demonstrates that the extents of DNase I digestion were similar for all reactions. Nucleotide numbering at the left is from the labeled end and corresponds to the published sequence (12). Asterisks represent the region of weak protection (see Results and Discussion). The locations of DNase I protection regions are indicated at the right by brackets. For IRI, only the right arm of the repeat is shown since the region corresponding to the left arm is undetectable under the conditions of the footprint. L and R designate the left and right arms, respectively, of the repeats. Locations of the inverted repeats are shown by arrows. Quantities of purified BtgA in reaction mixtures: lane C, none lanes 1 to 5, 100 ng, 300 ng, 500 ng, 1 μg, and 3 μg, respectively. (A) Cleavage pattern of the lower strand of oriT. The BtgA- oriT complexes were subjected to limited cleavage by DNase I and electrophoresed on 8% polyacrylamide sequencing gels. ![]() The specifically labeled upper and lower strands of oriT were incubated with increasing amounts of purified BtgA protein in binding reactions. DNase I footprint analysis demonstrated that BtgA binds apparently in a single-stranded fashion to the oriT-containing fragment, overlapping inverted repeats I, II, and III and the putative nick site.ĭNase I footprint analysis of the BtgA-pBFTM10 oriT protein complex. coli, and the purified protein was used in electrophoretic mobility shift assays, demonstrating specific binding of BtgA protein to its cognate oriT. A 304-bp DNA fragment comprising this putative oriT region was cloned and confirmed to be the functional pBFTM10 oriT by bacterial conjugation experiments using E. DNA sequence analysis of the region immediately upstream of btgA revealed three sets of inverted repeats, potentially locating the oriT region. The BtgA protein was predicted to contain a helix-turn-helix motif, indicating possible DNA binding activity. The Bacteroides fragilis conjugal plasmid pBFTM10 contains two genes, btgA and btgB, and a putative oriT region necessary for transfer in Bacteroides fragilis and Escherichia coli. ![]()
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