Escherichia coli cells transformed with pbr322 plasmids have a selective advantage in antibiotictreated environments. We used the pbr322 dna plasmid as a vector for the gene called cat chloramphenicol acetyltransferase, coding for antibiotic resistance to chloramphenicol. Tdna, from ti or ri plasmid of agrobacterium, is considered to be a very potential vector for cloning experiments with higher plants. Among higher plants, ti plasmid of agrobacterium tumefaciens and ri plasmid of. A cloning vector is a genome that can accept the target dna and increase the number of copies through its own autonomous replication. The replication origin of pbr322, which directs multiplication of the cloning vector in host cells is originally from pmb1. Finally a plasmid miniprep was used to isolate pbr322 from the e. This plasmid vector has been put together from fragments originating from three different naturally. The gene for chloramphenicol resistance cm has also been introduced. The molecule is a doublestranded circle 4,361 base pairs in length 2. Cloning vectors usually are selected on the basis of differences in their capacity for the. These fragments can serve as accurate dna size markers from small pieces up to the 4362 base pair length of pbr322. This file contains additional information such as exif metadata which may have been added by the digital camera, scanner, or software program used to create or digitize it. The rop gene product, which regulates plasmid replication by stabilizing the interaction between rnai and rnaii transcripts, maintains the copy number at about 20 per cell.
Welcome to vector database vector database is a digital collection of vector backbones assembled from publications and commercially available sources. Pdf pbr322 vectors having tetracyclinedependent replication. Vector pam34, the iptgdependent parental plasmid was also included in these analyses. Another plasmid used as a vector to clone dna is called puc18 plasmid. Psfcmvpuc18 cmv puc18 mcs plasmid plasmid vector for molecular cloning. If the file has been modified from its original state, some details such as the timestamp may not fully reflect those of. As a starting material for cloning of casein gene sequences. The plasmid pbr322 was one of the first ek2 multipurpose cloning vectors to be designed and constructed ten years ago for the efficient cloning and selection of recombinant dna molecules in.
Information from its description page there is shown below. Get a printable copy pdf file of the complete article. The effects of ampicillin versus tetracycline on the. First, the vector dna and cat dna was cut with the. The pbr322 vector 4,361bp carries the genes for tetracycline and ampicillin resistance. The phenotype of bacteria containing the intact plasmid is amp r tet r. Vector pbr322 this artificial vector was constructed by bolivar and rodriguez in 1977. This page is informational only this vector is not available from addgene please contact the manufacturer for further details. Promega corporation spend more, get more on promega products. Cloning vectors cloning vectors are dna molecules that are used to transport cloned sequences between biological hosts and the test tube. Pdf the plasmid pbr322 was one of the first ek2 multipurpose cloning vectors to be designed and constructed ten years ago for the. Promoter p2 in the same region as p1, but it is on the opposite strand and initiates transcription in the direction of the tetracycline resistance gene. To convert pdf to vector format, it is necessary to convert a pdf to bitmap image firstly and then you can easily convert the images to vectors.
Jenny solloway section k gel electrophoresis is useful for separating dna by size and the information that can be inferred from the data obtained from a dna gel. By default, macvector assumes that new sequences have a linear topology. Inserts cloned in this vector may be characterized easily using this data. Biology 335 lecture notes recdna plasmid vectors i. The amp r gene originally residing on plasmid r1, tet r from r65. This plasmid contains a gene that makes the host cell ampicillin resistant. Because pbr322 is a circular molecule, the first thing we must do is tell macvector that it is actually a circular molecule. While the file is a simple text file, macvector automatically interprets the contents and works out that it must be a dna sequence. Pdf plasmid vector pbr322 and its specialpurpose derivativesa.
The enzyme produces a blue pigment when exposed to a specific substrate analog. The pbr322 plasmid is a commonly used cloning vector that contains both the ampicillin and tetracycline resistance genes as selectable markers. This is a free resource for the scientific community that is compiled by addgene this page is informational only this vector is not available from addgene please contact the manufacturer for further details. In order to use antibiotic resistance as a negative as well as a positive selection system, the plasmid vector must carry two different antibiotic resistance genes. It is approximately 4300 bp in length and has two antibiotic resistance genes. Hipdf is a tool that will help users convert pdf to various file formats, vice versa. Thermo scientific pbr322 is one of the most commonly used li cloning vectors.
Vector database is a digital collection of vector backbones assembled from publications and commercially available sources. Plasmid vector pbr322 and its specialpurpose derivatives. Results from a previous experiment indicate that different selective. The maps, notes, and annotations in the zip file on this page are ed material. Created in 1977 in the laboratory of herbert boyer at the university of california, san francisco, it was named after francisco bolivar zapata, the postdoctoral researcher who constructed it. The attached sequence file has been compiled from information in the sequence databases, published literature, and other sources, together with partial sequences obtained by clontech laboratories, inc. The plasmid contents of these strains were compared to that of pbad322s, a 5535 bp spectinomycindependent vector that carries the wild type pbr322 origin and is of a size quite similar to that of pcy1107 cronan, 2006. Promoter p3 is the natural promoter for the betalactamase gene. The p stands for plasmid, and br for bolivar and rodriguez. Promoter p1 is artificially created by the ligation of two different dna fragments to create pbr322. Structural biochemistrydna recombinant techniquesplasmid.
Ampicillin versus tetracycline in the selection of pbr322. It can be a plasmid, a bacteriophage, or yeast artificial chromosome yac. Plasmids pbr322 and puc8 pbr322 plasmid one of the first plasmids to be used in recombinant genetics was called pbr322. The plasmid pbr322 was one of the fast ek2 mtipose cloning vectors to be designed and constructed. It also contains a gene that allows it produce betagalactosidase, which is an enzyme degrades certain sugars. It is one of the most popular and commercially used early purpose built plasmid based vector. This material may be used without restriction by academic, nonprofit, and governmental entities, except that the source must be cited as. Recombinant dna pancasila pdf pbr322 cloning vehicle messenger rna purification hybridizations. Principles of gene manipulation and genomics pdf 7th ed. These combined dna sequence and map files can be opened with snapgene or the free snapgene viewer. Pelley phd, in elseviers integrated biochemistry, 2007.
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