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Original link:Metaorganisms - Plasmid Construction
The Heyuan Biological Research Service Platform can provide cutting-edge, comprehensive, and rigorous vector design and construction services for gene function researchers, covering ordinary eukaryotic expression vectors, viral vectors, and more.
The virus vector construction service includes lentivirus, adenovirus, adenovirus, and retrovirus vectors, etc. The construction content includes gene overexpression vector construction, gene directed mutation expression vector construction, shRNA design and construction for gene silencing, microRNA expression and blocking vector design and construction, luciferase reporter gene vector construction (UTR, promoter, and other functional sequences), as well as Cre LoxP system and CRISPR/Cas9 gene editing tool vector design and construction, aiming to solve the needs of gene upregulation and gene downregulation in gene function research in a one-stop manner.
Plasmid advantages:
① The expression time is fast, and the target gene expression can be observed about 24 hours after being introduced into the cell;
② The construction technology is simple and the cost is low.
③ The overexpression vector library of Heyuan is abundant, and the overexpression markers are abundant
Fluorescent labeling：GFP、mCherry、CFP、BFP、EYFP、mNeonGreen、tdTomato、 mScarlet…
Label marking: FLAG、 HA 、Myc、His tag…
Resistance markers：Puro、Neo、Hygro、BSD…
limitations：
① Transfection reagents have cytotoxicity and can affect cell status after use;
② Most cells are difficult to transfect and cannot achieve genetic manipulation goals;
③ Plasmids are not integrated into the cell genome and can only be expressed instantaneously for a short period of time, making it impossible to conduct experiments with longer observation cycles.
Due to the limitations of pure plasmids, they need to be packaged into viruses to further meet the needs of different experiments.
Differences in different expression vectors
expression system | eukaryotic expression vector | Lentiviral vector | Recombinant adenovirus vector | Recombinant adenovirus vector |
Carrier gene characteristics | Double stranded DNA plasmid | RNA retrovirus | Single stranded DNA virus | Double stranded DNA virus |
Exogenous gene expression time | Expression starts at 24 hours and lasts for 3-7 days | Expression starts from 2-4 days and stabilizes over a long period of time | Starting expression from 7-14 days | Starting from 1-2 days to express |
Insert fragment size | Around 8kb | About 4kb | About 2.8kb | About 6kb |
Stable cell line screening | Difficult to operate | sure | may not | No, it can't be expressed instantaneously |
Cell experiments | Cell lines with low transfection efficiency | First choice, broad-spectrum, high infection efficiency | unsuited | Broad spectrum, high infection efficiency |
Animal experiments | unsuited | Suitable, depending on observation time and injection site | First choice, based on observation time and injection site | High immunogenicity, careful selection |
Titer range | nothing | 108~109TU/ml | 1012-1013vg/ml | 1010-1011pfu/ml |
