Molecular Cloning
overview

Create a DNA vector construct of your choice with us. From RNA interference and GFP-marked proteins to complex fusion proteins, we offer comprehensive assistance. Our services within molecular cloning range from identifying the most effective organism-specific promoter and selection marker, providing advanced 2D and 3D modeling to fine-tune the expression product and whole gene synthesis.

 

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Tailored vector backbone elements: Having a comprehensive database of vector backbone elements before designing a complicated DNA vector is critical. These elements - including a range of promoters, expression control sequences, transcription terminators, protein linkers, and protein tags - each play a crucial role in the gene expression process, affecting the efficiency, localization, and control of the output. Access to a vast selection of these elements allows us to tailor the vector design to the specific needs of the project, optimizing for factors like expression strength, stability, or regulatory control.

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Gene expression validation: Gene expression process is influenced by many factors including the promoter, terminator, and vector backbone used, all of which can affect the efficiency of gene expression. Each of these elements may interact differently with different genes or host organisms, leading to variability in expression levels. Therefore, verifying the expression level post-cloning is crucial to ensure the vector is functioning as intended and that sufficient protein or RNA is being produced for downstream applications.

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3D structure prediction: Designing fusion proteins that retain their full functionality and biological activity is a complex task, and this is where our cutting-edge 3D structure prediction service steps in. This process is essential to ensure the proteins you're studying maintain their structure and function, even when fused with additional tags or proteins.

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Ready to use full materials and methods and publication data: Providing a ready-to-use, comprehensive materials and methods and descriptions is a key component of our service. Drafting a materials and methods section can be time-consuming and complex, potentially requiring several hours to days of a researcher's time depending on the complexity of the procedures used. By supplying a detailed, publication-ready description, we free up significant time for our customers.

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Examples Of What You Can Do

Designing a DNA vector construct to successfully express a specific recombinant protein or RNA can be challenging, particularly for researchers unfamiliar with this specialized process. Numerous elements contribute to these complexities. For instance, there’s a vast array of transcription promoters available, yet not all yield efficient results with the desired protein. An incorrect promoter choice can result in low or even non-existent protein expression.

 

Similarly, selecting the appropriate transcription terminator sequence can be daunting. While an array of transcription terminator sequences exists, not all function optimally with the given promoter, protein, or vector backbone. An ill-suited terminator can trigger elongated or unstable RNA transcripts, leading to low protein expression.

 

Furthermore, the recombinant protein may originate from a different species, and its codons may not be optimized for the intended bacterial host. This mismatch can lead to sub-optimal protein expression levels.

 

These challenges can be bypassed by employing a professional service to design and assemble a function-specific, custom vector. Leveraging our expertise in molecular cloning, gene synthesis, and bioinformatics, we ensure optimal vector design and assembly for successful recombinant protein or RNA production. Our services are geared towards facilitating efficient gene expression and streamlining your research process, regardless of the complexity of your target protein or RNA.

Understanding the activity and localization of proteins can be achieved by fusing the protein of interest with a reporter protein, such as the commonly utilized Green Fluorescent Protein (GFP). When this fusion occurs, GFP will fluoresce, enabling researchers to track the location and activity of the protein in real-time. This monitoring allows in-depth insight into the protein’s function and regulation.

 

Alternatively, fusing a protein of interest with a mitochondrial targeting sequence or a Nuclear Localization Signal (NLS) can facilitate the relocation of a specific protein to a designated organelle. However, this process often encounters challenges as most fusion proteins fail to function effectively. The functionality of the native protein can be significantly impaired by the protein or proteins attached to it.

 

To overcome these hurdles, the 3D structure of the protein needs to be considered, and a suitable protein linker sequence – from the over 100 available – must be carefully selected for the task at hand. At Deep Biotech Solutions, our specialists in 3D structure prediction, combined with our proprietary fusion linker peptides, offer tailored solutions to these challenges. By leveraging our expertise in molecular cloning, bioinformatics, and artificial intelligence, we ensure optimal fusion protein function for accurate investigation of protein activity and localization.

RNA interference (RNAi) offers powerful research potential by enabling the selective silencing of specific genes across a range of organisms, from plants and animals to humans. RNAi can be utilized to study gene function, identify prospective therapeutic targets, and pioneer novel treatments for diseases.

 

Despite its potential, designing and constructing an efficient RNAi vector can present significant challenges. A key issue is ensuring the RNAi vector specifically targets the gene of interest, operating with high efficiency without inadvertently impacting other genes. The effectiveness and precision of RNAi largely depend on the choice of transcriptional promoter, terminator, vector backbone, and sequence of the short interfering RNA (siRNA) employed to target the gene.

 

To avoid off-target effects such as unintentional silencing of similar sequence genes, the siRNA must be meticulously designed. Moreover, the choice of the RNAi vector is contingent upon the specific experimental design and the organism under study. Some organisms may necessitate specialized RNAi vectors to achieve successful gene silencing.

 

At our company, we bring the advantage of expert guidance in vector compilation. Our team of bioinformaticians and geneticists work together to optimize every aspect of the vector, utilizing personalized gene synthesis to custom-fit the ordered vector construction to your specific task. Our services in NGS sequencing, molecular cloning, bioinformatics, artificial intelligence, and gene synthesis ensure the delivery of high-quality, task-specific RNAi vectors.

Fusion proteins are hybrid entities created by conjoining two or more protein domains, each possessing a unique function. This fusion process often results in a protein with enhanced or completely new properties, with widespread applications in both scientific research and various industries.

 

Below are a few examples demonstrating the utility of fusion proteins with newly attributed functions:

 

  • Antibody Fusion Proteins: These are created by fusing an antibody domain with another protein domain, such as a toxin or an enzyme. The resulting antibody fusion proteins can specifically target and destroy cancer cells or viruses.
  • Enzyme Fusion Proteins: These are generated by fusing an enzyme domain with a binding domain, such as an antibody. Enzyme fusion proteins can selectively bind to and catalyze reactions on specific targets, including pathogens, different polymers, plastics, or other waste products.
  • Reporter Fusion Proteins: Created by fusing a reporter domain with a protein of interest, these fusion proteins facilitate real-time monitoring of protein activity and localization in living cells. Even complex fusion proteins designed for specific functions, such as those containing a quadruple fusion to monitor autophagic flux (GFP-LC3-RFP-LC3ΔG), can be produced.
  • Purification Fusion Proteins: These are produced by fusing a purification domain, such as a histidine tag or a streptavidin-binding domain, with a protein of interest. Purification fusion proteins enable efficient and selective purification of the protein.

 

The development of fusion proteins begins with an accurate 3D conformational design or involves trial and error with various arrangements and protein linkers until a functional fusion protein is achieved. At Deep Biotech Solutions, our team of 3D structure prediction specialists and optimized fusion linker peptides, developed in-house, can assist with these challenges. Our services in NGS sequencing, molecular cloning, bioinformatics, artificial intelligence, and gene synthesis ensure the production of high-quality, custom-fit fusion proteins for your specific research needs.

Frequently Asked Questions

Molecular cloning is a process used to create identical copies (or clones) of a specific DNA segment. This enables researchers to study the function of specific genes and proteins.

We offer comprehensive assistance, from creating a DNA vector construct of your choice to complex tasks such as RNA interference and Green Fluorescent Protein (GFP)-marked proteins. We can also handle advanced 2D and 3D modeling to fine-tune the expression product (RNA or protein) and conduct whole gene synthesis.

Molecular cloning has numerous applications, including gene function studies, gene therapy, production of recombinant proteins, and in the development of genetically modified organisms.

Gene synthesis is a method in synthetic biology that creates specific, custom DNA sequences. This synthesized DNA can then be cloned into a DNA vector for further study or manipulation.

Advanced 2D and 3D modeling helps to predict how the protein or RNA coded by your sequence of interest will behave. This can provide crucial insights into protein function and interactions or RNAi efficiency.

GFP-marked proteins enable real-time visualization of protein location and behavior within a cell, providing valuable insights into protein function and cellular processes.

Vector NGS validation confirms the correct sequence of the cloned gene and ensures there are no unintended mutations, ensuring the reliability of your research outcomes.

Yes, we can create a custom DNA vector construct tailored to your specific research needs, enabling precise control over your genetic experiments.

Reach out to us if you have any queries about our products. We assure you of a response within 24 hours.

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