Structure of inserted bacteriophage Mu-1 DNA and physical mapping of bacterial genes by Mu-1 DNA insertion.

نویسندگان

  • M T Hsu
  • N Davidson
چکیده

It is shown, by electron microscope observation of the structures of heteroduplexes, that Mu-1 DNA inserted into the bacterial episomes Flac and F(8)[1] is collinear with, rather than a circulation permutation of, the DNA of the mature Mu-1 bacteriophage. Observation of the position of the inserted Mu defines a point within the gene that has been inactivated (the lacI gene for Flac and a transfer gene in F(8)[1] in these particular instances). These examples illustrate a new, general method for physical gene mapping. The episome with Mu DNA inserted into F(8)[1] [i.e., F(8)[1](Mu)], although derived from a single colony, is heterogeneous in that a self-renatured sample shows a nonhomology loop of length 3.0 kb. This nonhomology loop, which has previously been observed in mature Mu-1 DNA, is due to an inversion.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Efficient insertion mutagenesis strategy for bacterial genomes involving electroporation of in vitro-assembled DNA transposition complexes of bacteriophage mu.

An efficient insertion mutagenesis strategy for bacterial genomes based on the phage Mu DNA transposition reaction was developed. Incubation of MuA transposase protein with artificial mini-Mu transposon DNA in the absence of divalent cations in vitro resulted in stable but inactive Mu DNA transposition complexes, or transpososomes. Following delivery into bacterial cells by electroporation, the...

متن کامل

Mini-mu bacteriophage with plasmid replicons for in vivo cloning and lac gene fusing.

New mini-Mu transposons with plasmid replicons were constructed with additional features for in vivo DNA cloning and lac gene fusing in Escherichia coli. These mini-Mu replicons can be used to clone DNA by growing them with a complementing Mu bacteriophage and by using the resulting lysate to transduce Mu-lysogenic cells. These mini-Mu phage have selectable genes for resistance to kanamycin, ch...

متن کامل

Complete genomic sequence of bacteriophage B3, a Mu-like phage of Pseudomonas aeruginosa.

Bacteriophage B3 is a transposable phage of Pseudomonas aeruginosa. In this report, we present the complete DNA sequence and annotation of the B3 genome. DNA sequence analysis revealed that the B3 genome is 38,439 bp long with a G+C content of 63.3%. The genome contains 59 proposed open reading frames (ORFs) organized into at least three operons. Of these ORFs, the predicted proteins from 41 OR...

متن کامل

The role of MuB in selecting transposition targets of bacteriophage

Phage Mu transposes promiscuously, employing MuB protein for target capture. MuB forms stable filaments on A/T-rich DNA, and a correlation between preferred MuB binding and Mu integration has been observed. We have investigated the relationship between MuB-binding and Mu insertion into ‘hot’ and ‘cold’ Mu targets within the E. coli genome. Although higher binding of MuB to select hot versus col...

متن کامل

The regulatory region of the L-arabinose operon: a physical, genetic and physiological study.

A f&-structure physical and genetic m&p wt~s constructed of a 1000 base-pair region of t,he L-arabinose operon of Escherichia coEi. This region consists of the ara regulatory sequences contained between the araC and araB genes and portions of these flanking genes. Point mutations, Mu phage insertions, and bacterial deletions 8s well as arabinose-induced and basal enzyme levels in the strains we...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:
  • Proceedings of the National Academy of Sciences of the United States of America

دوره 69 10  شماره 

صفحات  -

تاریخ انتشار 1972