What are Sf9 cells used for?

Sf9 cells, a clonal isolate of Spodoptera frugiperda Sf21 cells (IPLB-Sf21-AE), are commonly used in insect cell culture for recombinant protein production using baculovirus. They were originally established from ovarian tissue. They can be grown in the absence of serum, and can be cultured attached or in suspension.

How do you transfect Sf9 cells?

Warm Transfection Medium to room temperature 2. Transfer 2.5mL of Sf9 at 1 x 10 cells/mL into each 24 deep well block. Add 3 uL FuGENE HD directly to the Transfection Medium, avoid touching the side of the tube when adding FuGENE. Incubate at room temperature for 5 minutes.

How does baculovirus expression system work?

In baculovirus expression system, cotransfection of AcNPV (Autographa californica nuclear polyhedrosis virus) DNA with a compatible recombinant transfer plasmids containing coding sequence is done in insect cells. Establish the cell line, Sf9 or Sf21, in Transfection medium (Expression systems #95-020).

What is an insect cell expression system?

Insect cells offer high levels of protein expression with posttranslational modification approaching that of mammalian cells, ease of scale-up, and simplified cell growth that can be readily adapted to high-density suspension culture for large-scale expression.

What are CHO cells and what are they used for?

CHO cells are the most common mammalian cell line used for mass production of therapeutic proteins. They can produce recombinant protein on the scale of 3-10 grams per liter of culture.

What is a Bacmid?

The bacmid is essentially a very large plasmid containing the baculovirus genome modified to carry a lacZ gene and an attTn7 docking site inserted in the lacZ coding region. The most commonly used cell line for expressing recombinant proteins from baculovirus vectors is Sf9.

What is Bacmid DNA?

The bacmid is essentially a very large plasmid containing the baculovirus genome modified to carry a lacZ gene and an attTn7 docking site inserted in the lacZ coding region. The helper plasmid expresses the Tn7 transposase.

Can baculovirus infect human cells?

These results show that baculovirus can infect nondividing mammalian cells, which implies that the viral genome is able to penetrate the mammalian nuclear membrane.

What is an expression system?

Definition. Expression systems are genetic constructs (a gene encoded by DNA) that are designed to produce a protein, or an RNA (ribonucleic acid), either inside or outside a cell. Expression systems are used in research and in the commercial production of enzymes or therapeutics.

What is the advantage of the yeast expression system?

Yeast is a eukaryotic organism and has some advantages and disadvantages over E. coli. One of the major advantages is that yeast cultures can be grown to very high densities, which makes them especially useful for the production of isotope labeled protein for NMR.