What is the size of pbluescript?

Plasmid: pBluescript SK (+)

Source/Vendor: Stratagene
Plasmid Type: Bacterial Expression
Cloning Method: Restriction Enzyme
Size: 2958
5′ Sequencing 1 Primer: M13pUC-fwd

What is pBluescript SK+?

The pBluescript II SK+ phagemid (plasmids with a phage origin) is a cloning vectors designed to simplify commonly used cloning and sequencing procedures, including the construction of nested deletions for DNA sequencing, generation of RNA transcripts in vitro and site-specific mutagenesis and gene mapping.

Is pBluescript a restriction enzyme?

The pBluescript II phagemids have an extensive polylinker with 21 unique restriction enzyme recognition sites. Flanking the polylinker are T7 and T3 RNA polymerase promoters that can be used to synthesize RNA in vitro.

Why is pBluescript a good vector?

The versatility of pBluescript vectors alleviates the need for successive subcloning. With 21 restriction sites, in two orientations, most cloned genes can be inserted directionally into the pBluescript polylinkers. … If a new gene of unknown sequence is to be cloned, libraries will need to be generated for screening.

What is polylinker sequence?

A polylinker is a short DNA sequence containing two or more different sites for cleavage by restriction enzymes. Polylinkers are introduced into vectors to make cloning easier by providing sites that allow cloning DNA, cut with any of a number of different restriction enzymes, into a single plasmid.

What is pBluescript II?

The pBluescript II phagemids (plasmids with a phage origin) are cloning vectors designed to simplify commonly used cloning and sequencing procedures, including the construction of nested deletions for DNA sequencing, generation of RNA transcripts in vitro and site-specific mutagenesis and gene mapping.

What is F1 origin?

The ori is the place where DNA replication begins, enabling a plasmid to reproduce itself as it must to survive within cells (Addgene). F1 is a phage-derived ori that allows for the replication and packaging of ssDNA into phage particles. Plasmids with phage-derived ori’s are referred to as phagemids.

What is pUC18 plasmid?

$ 40.00. pUC18 is a commonly used plasmid cloning vector in E. coli. The molecule is a double-stranded circular DNA (2686 base pairs in length). Due to a small size pUC18 enables successful cloning of large DNA fragments.

What is a phagemid vector?

A phagemid or phasmid is a DNA-based cloning vector, which has both bacteriophage and plasmid properties. These vectors carry, in addition to the origin of plasmid replication, an origin of replication derived from bacteriophage.

Do plasmids replicate?

The plasmid is a small DNA molecule within a chamber that is physically separated from chromosomal DNA and can replicate independently [6].

What is shuttle vector example?

A vector that can replicate in more than one host organisms or two different cell types (e.g. a prokaryotic cell and a eukaryotic cell). An example is the yeast shuttle vector that can propagate within the cells of E. coli and yeast.

What is pBin19?

Plasmid: pBin19

(ATCC staff) A vector for efficient transformation of higher plants in a binary plasmid system with pAL440. (personal communication) Medium is 1065 LB medium. NCBI gi: 520486 NCBI gi: 529322 Hosts: Agrobacterium tumefaciens, E. … Binary Agrobacterium vector for plant transformation.

What does the M13 fragment in a phagemid contain?

3. What does the M13 fragment in a phagemid contain? Explanation: These signal sequences are recognized by the enzymes that convert the normal double-stranded M13 molecule into single-stranded DNA before secretion of new phage particles. 4.

What are polylinker for plasmids?

A multiple cloning site (MCS), also called a polylinker, is a short segment of DNA which contains many (up to ~20) restriction sites – a standard feature of engineered plasmids. … The purpose of an MCS in a plasmid is to allow a piece of DNA to be inserted into that region.

What is insertional activation?

Definition. Insertional activation/inactivation refers to either activation of an endogenous gene which is located near an integrated transgene, or to disruption of a gene or other functional sequence by insertion of a transposable element.

What is MCS of a plasmid?

Multiple Cloning Site (MCS) Short segment of DNA which contains several restriction sites allowing for the easy insertion of DNA. In expression plasmids, the MCS is often downstream from a promoter.

What do restriction enzymes do in bacteria?

A restriction enzyme is an enzyme isolated from bacteria that cuts DNA molecules at specific sequences. The isolation of these enzymes was critical to the development of recombinant DNA (rDNA) technology and genetic engineering.

What is the difference between cloning and expression vectors?

Cloning vectors are the DNA molecules that carry a specific gene of interest into the host cell and its main purpose is to make numerous copies of the inserted gene. … Expression vectors are associated with the actual expression of the gene into mRNA and protein in the target organism.

What is the purpose of using restriction enzymes on a purified plasmid?

When cloning by restriction digest and ligation, you use restriction enzymes to cut open a plasmid (backbone) and insert a linear fragment of DNA (insert) that has been cut by compatible restriction enzymes.

How fast is an F1 car?

Formula 1 cars have a top speed of 360KPH (223MPH) but have been known to reach speeds closer to 400KPH (248MPH). These cars accelerate from 0 – 100KPH (62MPH) in 2.4 seconds and have cornering speeds of 300KPH (186MPH). There are no cars faster around a racetrack than F1 cars.

Who invented F1?

Formula One Group

Formerly Formula One Constructors Association
Type Subsidiary
Industry Motorsport
Founder Bernie Ecclestone
Area served Globally

Is F1 a car?

Formula One (also known as Formula 1 or F1) is the highest class of international racing for open-wheel single-seater formula racing cars sanctioned by the Fédération Internationale de l’Automobile (FIA).

What is the size of pUC18?

Thermo Scientific pUC18 vector is a small, high copy number, E. coli plasmid, 2686 bp in length. It contains identical multiple cloning site (MCS) as pUC19 vector except that it is arranged in opposite orientation.

How many base pairs is pUC18?

pUC18 is a commonly used plasmid cloning vector in E. coli.

The molecule is double-stranded circle, 2686 base pairs in length, and has a high copy number. pUC18 carries a 54 base-pair multiple cloning site polylinker that contains unique sites for 13 different hexanucleotide-specific restriction endonucleases.

How is pUC18 made?

pUC18 is a small, high copy cloning vector for replication in E. … It has been constructed using the ampicillin resistance gene and the pMB1 origin of replication from pBR322. The pMB1 of pUC18 differs from the pBR322 origin by a single point mutation and the lack of the rop gene, leading to a high copy number.

What is difference between phagemid and Phasmid?

As nouns the difference between phagemid and phasmid

is that phagemid is a phage whose genome contains a plasmid that can be removed during the infection of a host with a second, helper phage while phasmid is (entomology) any insect of the order phasmatodea, the leaf insects and walking sticks.

What is pUC19 plasmid?

pUC19 is a commonly used cloning vector that conveys the Amp resistance. The molecule is a small double-stranded circle, 2686 base pairs in length, and has a high copy number. … NEB offers a selection of common cloning plasmids and DNAs for use as substrates.

What is the difference between phagemid and plasmid?

is that plasmid is (cytology) a loop of double-stranded dna that is separate from and replicates independently of the chromosomes, most commonly found in bacteria]], but also in archaeans and [[eukaryote|eukaryotic cells, and used in genetic engineering as a vector for gene transfer while phagemid is a phage whose …

Why are plasmids small?

A plasmid is a small, often circular DNA molecule found in bacteria and other cells. Plasmids are separate from the bacterial chromosome and replicate independently of it. They generally carry only a small number of genes, notably some associated with antibiotic resistance.

How big is a plasmid?

The size of the plasmid varies from 1 to over 200 kbp, and the number of identical plasmids in a single cell can range anywhere from one to thousands under some circumstances.

What is vector copy number?

These vectors have the capacity to integrate permanently into host cell DNA. There is an increased risk of oncogenesis if the vector copy number (VCN) per cell is high. The Food and Drug Administration (FDA) recommends that the VCN shall be &lt,5 copies per genome.

Is Yep a shuttle vector?

Yeast episomal plasmids (YEps) are shuttle vectors. They can replicate in E. coli and also in yeast.

Why are shuttle vectors used?

Shuttle vectors are frequently used to quickly make multiple copies of the gene in E. coli (amplification). They can also be used for in vitro experiments and modifications (e.g. mutagenesis, PCR). One of the most common types of shuttle vectors is the yeast shuttle vector.

Is pBR322 a shuttle vector?

A series of shuttle vectors for Bacillus subtilis and Escherichia coli was developed. These are derived from one basic construct composed of parts of the Gram+ plasmid pUB110 and the Gram- plasmid pBR322. They contain multiple cloning sites flanked by transcriptional terminators.

What is PBIN19 Mcq?

Explanation: PBIN19 is a disarmed cloning vector which is a binary vector.

Which part of the T-DNA are involved in infection?

A key feature of Ti plasmids is their ability to drive the production of opines, which are derivatives of various amino acids or sugar phosphates, in host plant cells. These opines can then be used as a nutrient for the infecting bacteria, which catabolizes the respective opines using genes encoded in the Ti plasmid.

What is the basis of binary vector strategy Mcq?

14. What is the basis of “binary vector” strategy? Explanation: The binary vector strategy is based on the observation that the T-DNA does not need to be physically present/attached to the rest of the Ti plasmid. 15.

What is the size of M13 phage vector?

M13 phage is a bacterial virus composed of a single-stranded DNA encapsulated by various major and minor coat proteins. It has a long-rod filament shape that is approximately 880 nm in length and 6.6 nm in width (Smith &amp, Petrenko, 1997).

How is phagemid made?

phagemid A type of cloning vector developed as a hybrid of the filamentous phage M13 and plasmids to produce a vector that can grow as a plasmid, and also be packaged as single stranded DNA in viral particles.

What is a Fosmid library?

Fosmids are DNA vectors that use the F-plasmid origin of replication and partitioning mechanisms to allow cloning of large DNA fragments. A library that provides 20–70-fold redundant coverage of the genome can easily be prepared.

Where is the polylinker region?

A multiple cloning site (MCS, or Polylinker region) is a DNA region within a Plasmid that contains multiple unique Restriction enzyme cut sites. Plasmids are very useful in biotechnology and one key feature of their use is the multiple cloning site, which allows for foreign DNA to be inserted into the plasmid.

What is a polylinker quizlet?

A polylinker is a small region of DNA that contains multiple restriction sites, not found in other places of the plasmid.

What is incorrect plasmid?

What is incorrect about plasmid? Explanation: Plasmids are the extra genetic materials that are found in the bacterial cell along with the genetic component. They are autonomously replicating cyclic double strand DNA molecules used as vectors for gene transfer and also for replication.

What is the insertional inactivation?

Insertional inactivation technique of recombinant DNA technology used to select bacteria that carry recombinant plasmids, a fragment of foreign DNA is inserted into a restriction site within a gene for antibiotic resistance, thus causing that gene to become nonfunctional.

Why are white colonies desirable?

If a bacterium picked-up the original plasmid containing the intact B-galactosidase gene, they will hydrolyze X-gal to produce blue-colored colonies. 6. Therefore, white colonies are the desirable ones.

How is insertional activation and blue white screening applied?

One of the most influential selection methods of recombinant plasmid for the insertional inactivation procedure is a method known as ‘Blue-white’ selection method. In this procedure, the lacZ gene which is a reporter gene is inserted in the vector.

How do you calculate MCS on a plasmid?

Multiple cloning site (MCS) – YouTube

What is cloning plasmid?

DNA cloning is the process of making multiple, identical copies of a particular piece of DNA. … A circular piece of plasmid DNA has overhangs on its ends that match those of a gene fragment. The plasmid and gene fragment are joined together to produce a gene-containing plasmid.

What is a plasmid Multicloning site quizlet?

What is a plasmid multicloning site? Provides multiple unique restriction sites for inserting DNA fragments into the cloning vector. Plasmids and other vectors that contain two origins of replication recognized by different host organisms are called vectors.