What is blasticidin resistance?

Blasticidin S is a microbial antibiotic that inhibits protein synthesis in both prokaryotes and eukaryotes. The blasticidin S-resistance gene (bsr), isolated from Bacillus cereus K55-S1 strain, was inserted into pSV2 plasmid vector and introduced into cultured mammalian cells by transfection.

How does blasticidin resistance work?

Blasticidin prevents the growth of both eukaryotic and prokaryotic cells. It works by inhibiting termination step of translation and peptide bond formation (to lesser extent) by the ribosome. This means that cells can no longer produce new proteins through translation of mRNA.

What is the role of antibiotic resistance gene in a vector?

Adding an antibiotic resistance gene to the plasmid solves both problems at once – it allows a scientist to easily detect plasmid-containing bacteria when the cells are grown on selective media, and provides those bacteria with a pressure to keep your plasmid.

What is BSD in plasmid?

Blasticidin S Deaminase Gene (BSD): a New Selection Marker Gene for Transformation.

How much Blasticidin should I take for selection?

The Blasticidin concentration required for selection in mammalian cells varies depending on the cell line used. Use 2-10 μg/ml Blasticidin for selection in mammalian cells.

How does geneticin work?

G418 (Geneticin) is an aminoglycoside antibiotic similar in structure to gentamicin B1. G418 blocks polypeptide synthesis by inhibiting the elongation step in both prokaryotic and eukaryotic cells. Resistance to G418 is conferred by the neo gene from Tn5 encoding an aminoglycoside 3′-phosphotransferase, APT 3′ II.

What are the properties of a good vector?

Properties of an ideal vector

• It should be replicate autonomously.
• A vector should be less than 10 KB in size.
• It should be easily isolated and purify.
• It should be easily introduced into the host cell.
• It should have suitable marker genes.

What is antibiotic resistance genes?

Resistance genes have emerged against almost all classes of antibiotics, even against those considered last resort. Mobile antibiotic resistance genes (ARGs) are associated with a variety of mobile genetic elements (MGEs) that enable these genes to spread to new hosts, even across taxonomic boundaries.

How much puromycin should I take for selection?

Puromycin antibiotic ensures effective positive selection of cells expressing the puromycin-N-acetyl- transferase (pac) gene. In mammalian cells, the recommended working concentration range for puromycin is 0.5 – 10 µg/ml.

What is G418 selection?

G418 antibiotic ensures effective positive selection for cells expressing the neomycin resistance (neo) gene. In mammalian cells, the recommended working concentration range for G418 is 0.1 – 2.0 mg/ml. Different cell types and cell culture conditions may require different concentrations of selection antibiotic.

What is the kanamycin resistance gene?

Kanamycin is used in molecular biology as a selective agent most commonly to isolate bacteria (e.g., E. coli) which have taken up genes (e.g., of plasmids) coupled to a gene coding for kanamycin resistance (primarily Neomycin phosphotransferase II [NPT II/Neo]).

How is blasticidin resistance conferred on a cell?

Resistance to blasticidin is also conferred by the blastcidin S acetyltransferase gene (bls) from Streptoverticillum sp, and the blasticidin S deaminase gene (BSD) from Apergillus terreus. Typically, mammalian cells are sensitive to blasticidin concentrations of 1-10 µg/ml, and bacteria to 25-100 µg/ml.

How is blasticidin used in a bsdcassette vector?

Blasticidin is a potent selection agent that allows selection of stable mammalian cell lines in as little as one week. The BsdCassette™ vectors simplify the construction of custom vectors that use Blasticidin resistance as a selectable marker. Each BsdCassette™ vector is designed for use in a specific host system and offers:

How is blasticidin resistance conferred on Bacillus cereus?

Resistance to blasticidin is conferred by the blasticidin resistance gene from Bacillus cereus (bsr), which codes for blasticidin-S deaminase [1]. Resistance to blasticidin is also conferred by the blastcidin S acetyltransferase gene (bls) from Streptoverticillum sp [2], and the blasticidin S deaminase gene (BSD) from Apergillus terreus [3].

What kind of gene is blasticidin’s deaminase?

Blasticidin S deaminase gene from Aspergillus terreus (BSD): a new drug resistance gene for transfection of mammalian cells. Biochim. Biophys.

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