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Lambda Phage based Genome Library Construction Service

Background Service Published Data FAQ Resources

Through years of dedicated research, Creative Biolabs has successfully developed and commercialized an integrated service portfolio based on phage display technology. Currently, Creative Biolabs offers a unique generation service to construct genome library for our global clients by using our novel lambda phage platform.

Background

Bacteriophage lambda is a temperate bacteriophage of E. coli, which can undergo either lytic or lysogenic lifecycle. In the lysogenic state, the phage is stably replicated along with the host by integrating its genome into the bacterial chromosome; while in the lytic state, progeny phages are rapidly assembled and released by cell lysis, just like T4 and T7. Lambda phage has an icosahedral head composed of two major coat proteins, gpE (415 copies) and gpD (405-420 copies), and a single tail composed of gpV. Inside the head there contains its double-strand DNA genome (48502 bp) with two cohesive ends.

In contrast to conventional filamentous phage vehicles, lambda phage offers unique advantages. Particularly, lambda phage represents a feasible choice for constructing complex genome libraries because of its simple structure and flexible DNA. A genomic library is a set of clones that collectively contain every DNA sequence in the genome of a specific organism. With the help of restriction enzymes, extracted genome DNA can be prepared as numerous fragments (15-20 kbp) and inserted into engineered lambda vectors. After packaging the ligated recombinant DNA into lambda phage particles and amplification in host bacterial, a comprehensive library of phage clones each carry a different DNA fragment can be obtained, which collectively constitute the whole genome sequence of the indicated organism.

Lambda phage genomic library contributes to various applications, especially for antigen discovery and immune response investigations. By constructing genome libraries derived from specific pathogens (i.e. virus, bacteria or organ) and screening against the whole antibody repertoire of infected individuals, efficient identification of a large panel of antigenic regions can be achieved. Therefore, lambda phage genome library provides a highlight for improving diagnosis, prophylaxis, and therapy for many life-threatening diseases.

Lambda Phage-based Genome Library Construction

In Creative Biolabs, we can offer multiple lambda vectors containing different restriction sites for your choice. We are able to generate high-quality genome libraries with a trace amount of starting materials, including genomic DNA, chromosomes, uncultured environmental microbes, etc. Special measures are taken to generate DNA fragments with considerate representativeness, which ensures over 95% coverage of the original genome. In addition, we also provide professional screening service against various probes or antibodies.

With our state-of-the-art technology platforms and extensive expertise in phage derived technologies, Creative Biolabs has confidence in delivering first-class lambda phage genome libraries in the most satisfactory manner. In addition, our scientists are also able to offer custom service of lambda phage cDNA library construction as an alternative.

Published Data

Fig. 1 Schematic representation of the promoter region of the LH beta gene from a library screening of the Moroccan sheep database sequence: Homology sequence analysis revealed that the two sequences shared more than 96% homology with the reference sequence available in the database after alignment.1

The study focuses on the identification and characterization of a novel promoter region in the ovine LH-beta gene using a lambda phage-based genomic library. The research enhances the understanding of the genetic regulation of the LH-beta gene, which plays a crucial role in reproductive performance in sheep. The study's results demonstrate the discovery of a 503 bp upstream promoter region that significantly increases promoter activity when combined with the known sequence, providing new insights into the genetic mechanisms underlying reproductive traits. The lambda phage-based genomic library allowed for the efficient cloning and screening of large genomic fragments, facilitating the identification of previously unknown regulatory elements. This highlights the importance of lambda phage-based libraries in uncovering novel genetic sequences that contribute to gene regulation and expression, which can have significant implications for animal breeding and genetic research.

Reference
  1. Aherrahrou, Redouane, et al. "Identification of a novel ovine LH-beta promoter region, which dramatically enhances its promoter activity." SpringerPlus 4 (2015): 1-9. Distributed under Open Access license CC BY 4.0, without modification.

FAQ

  1. What is a lambda phage-based genomic library, and how is it constructed?

    A lambda phage-based genomic library is a collection of DNA fragments from an organism's genome, cloned into lambda phage vectors. To construct it, genomic DNA is isolated and digested with restriction enzymes, creating fragments that are then inserted into the lambda phage vectors. The recombinant phages are packaged and propagated in bacterial hosts, allowing for the storage and screening of large genomic DNA sequences, which is particularly useful for identifying and analyzing genes and regulatory elements.

  2. What are the key advantages of using a lambda phage-based genomic library in genetic research?

    Lambda phage-based genomic libraries offer several advantages, including the ability to clone large DNA fragments (up to 20 kb), which is beneficial for capturing entire genes and their regulatory regions. The high cloning efficiency of lambda phage allows for the construction of comprehensive libraries, making it easier to identify rare sequences. Additionally, the well-established techniques for manipulating and screening lambda phage make these libraries a valuable tool for genetic mapping, gene discovery, and functional genomics studies.

  3. How is a lambda phage-based genomic library screened to identify specific genes or sequences?

    Screening a lambda phage-based genomic library typically involves hybridization techniques, where labeled probes complementary to the target gene or sequence are used to identify positive clones. The library is plated on bacterial lawns, and plaques containing recombinant phages are transferred to membranes. The membranes are then hybridized with the labeled probes, and positive signals are detected, allowing researchers to isolate the phage clones that contain the desired genomic fragments.

  4. What types of applications are lambda phage-based genomic libraries used for?

    Lambda phage-based genomic libraries are used in gene cloning, identification of regulatory elements, studying gene function, and creating physical maps of genomes. They are also employed in comparative genomics to identify conserved sequences between species and in the analysis of genetic variations that contribute to phenotypic traits, such as disease susceptibility or agricultural productivity.

  5. What are the limitations of lambda phage-based genomic libraries?

    Its one limitation is the size of the DNA fragments that can be cloned, which is typically up to 20 kb, making it challenging to capture very large genes or entire operons. Additionally, the process of constructing and screening these libraries can be time-consuming and labor-intensive. Furthermore, some genomic regions, such as those with repetitive sequences, may be underrepresented or difficult to clone in lambda phage vectors.

  6. How does a lambda phage-based genomic library differ from a plasmid-based genomic library?

    The primary difference between lambda phage-based and plasmid-based genomic libraries lies in the vector used to clone the DNA fragments. Lambda phage vectors can accommodate larger DNA inserts (up to 20 kb) compared to plasmid vectors, which typically carry inserts of 1-10 kb. This makes lambda phage-based libraries more suitable for cloning large genes and regulatory regions. Additionally, lambda phage libraries are often easier to screen due to the high efficiency of phage propagation and plaque formation, whereas plasmid libraries require colony screening.

  7. What are the steps involved in isolating a gene from a lambda phage-based genomic library?

    First, the library is screened using a probe specific to the gene of interest. Positive phage plaques are identified, isolated, and purified. The DNA from the recombinant phage is then extracted and further analyzed, often through restriction mapping and sequencing, to confirm the presence of the target gene. Once identified, the gene can be subcloned into a plasmid vector for further functional studies.

  8. Can lambda phage-based genomic libraries be used for studying gene expression?

    While lambda phage-based genomic libraries are primarily used for cloning and mapping genomic DNA, they can also be used to study gene expression by identifying regulatory elements, such as promoters and enhancers, that control gene transcription. By cloning genomic regions upstream of a reporter gene in a lambda phage vector, researchers can analyze the regulatory activity of these sequences in different conditions or tissues, providing insights into gene expression patterns.

  9. How does the lambda phage vector contribute to the stability and efficiency of a genomic library?

    The lambda phage vector contributes to the stability and efficiency of a genomic library by providing a high-capacity cloning system that ensures the stable propagation of large DNA fragments. The vector's packaging mechanism allows for efficient encapsidation and delivery of recombinant DNA into host bacteria, leading to high cloning efficiency. Additionally, lambda phage vectors minimize rearrangements and deletions of cloned DNA, preserving the integrity of the genomic library and making it a reliable tool for gene identification and analysis.

Resources

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