S2 Cell Line Construction Service

With the increasing frequency, Schneider's embryonic Drosophila cell line 2 (S2 cell line) has been used to stably express various recombinant mammalian proteins, using plasmid vector cells specially designed to express proteins in Drosophila. Therefore, the S2 based system has all the advantages of the SF 9 based system. The main additional advantage is that S2 cells can be used to produce stable cell lines under the control of strong inducible promoters. Creative BioMart's technology team had developed a protocol of ineducable S2 cell line construction service that contains Mtn or hsp70 promoter. For clients who are interested in our services, please see below for more information.

Technology Description

S2 Cell Line Construction Service Important residues for Hsp70 dimerization (Takakuwa, 2019)

Although a variety of promoters have been shown to be effective on Drosophila S2 cells, Drosophila metallothionein (Mtn) and heat shock protein 70 (hsp70) promoters are strong inducible promoters. The expression protocol we developed describes the induction of MTN and HSP70 promoters. They can be combined to co-transfect cells.

Selection of expression system
Available expression option	Mtn-regulated S2 expression system Hsp70-regulated constitutive expression system
Construction of expression vector	Deliverable
Gene optimization Subcloning design Synthetic gene Subcloning to vector	1. Gene synthesis report 2. Sequencing validation report 3. Cloning strain containing expression plasmid (optional) 4. Expression plasmid 5. Plasmid instructions
Analysis and identification of protein expression test
Transfection Induction Expression test Expression analysis and identification	1. Positive expression strains 2. Small test expression process report 3. Try to express and analyze the original pictures 4. Small test expression summary
Protein expression and purification
Protein affinity purification The purified protein is identified by WB	1. Protein purification process report 2. Images of protein purification 3. Proteins 4. Western blot (labeled antibody) 5. Protein fingerprint (optional) 6. Molecular weight of protein mass spectrometry (optional)
Scale up
Amplification Culture	1. Protein purity identification report 2. Finished protein (agreed price according to the results of expression test)

Features of Inducible S2 Expression System

S2 cells can easily accept multiple copies of plasmid DNA transfection (up to 500 copies), so the protein expression level is high.
Under the guidance of P transposon, the foreign gene can be integrated into the chromosome of recipient cells stably.
S2 cells can grow as high-density suspension cultures with minimal maintenance and equipment
Protein expression can be strictly regulated by Mtn and hsp70 promoters. The promoters are highly inducible and have no basic activity in S2 cells.
Using the S2 system, a large number of homologous recombinant proteins can be produced in a relatively short time and with few equipment or space.

Related Services

In addition to the inducible cell line construction service, we also offer the following related services.

Customers who need our assistance are more than welcome to contact us or submit an online inquiry. Our professional team will provide you with detailed guidance and solution ASAP.

Reference

1. Takakuwa, J. E., Nitika, Knighton, L. E., & Truman, A. W. (2019). Oligomerization of Hsp70: Current Perspectives on Regulation and Function. Frontiers in molecular biosciences, 6, 81. https://doi.org/10.3389/fmolb.2019.00081

CONTACT OUR TEAM

Services