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Taatacgactcactataggg

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Decoding the Mystery: A Deep Dive into "taatacgactcactataggg"



The seemingly random string of letters "taatacgactcactataggg" might appear insignificant at first glance. However, to those familiar with the language of molecular biology, this sequence represents a powerful and fundamental element within the complex world of genetics. This article aims to unravel the significance of this specific sequence, exploring its context, function, and broader implications within the field of biotechnology.

1. Understanding the Sequence: A Primer on DNA



The sequence "taatacgactcactataggg" is a segment of deoxyribonucleic acid (DNA), the fundamental building block of life. DNA is a double-stranded helix composed of nucleotides, each consisting of a deoxyribose sugar, a phosphate group, and one of four nitrogenous bases: adenine (A), thymine (T), guanine (G), and cytosine (C). The sequence we are examining represents a single strand of DNA, written using the standard convention of representing only one strand (the other strand is easily deduced due to base pairing: A with T, and G with C).

This particular sequence bears a striking resemblance to a known promoter sequence. Promoters are crucial regions of DNA that act as binding sites for RNA polymerase, the enzyme responsible for initiating transcription – the process of creating RNA from a DNA template. The RNA then serves as a blueprint for protein synthesis.

2. The T7 Promoter: A Workhorse in Biotechnology



The sequence "taatacgactcactataggg" is remarkably similar, if not identical depending on the specific variant, to the strong promoter sequence recognized by the T7 RNA polymerase. The T7 promoter is a bacteriophage (a virus that infects bacteria) promoter that has been extensively utilized in molecular biology labs worldwide. Its strength lies in its high efficiency in driving transcription.

This makes it incredibly useful for various biotechnological applications. For example, researchers often use the T7 promoter to control the expression of genes in bacteria. By placing the gene of interest downstream of the T7 promoter, they can precisely regulate the production of the corresponding protein. This control is particularly important in producing large quantities of specific proteins for research, pharmaceutical purposes, or industrial applications.

3. Applications of the T7 Promoter and its Associated Sequence



The T7 system's versatility is illustrated in its wide range of applications:

Protein Production: The T7 promoter is frequently used in expression vectors to produce recombinant proteins in bacteria like E. coli. This is crucial for manufacturing therapeutic proteins, enzymes for industrial processes, and proteins for structural and functional studies.

Gene Expression Studies: Researchers use the T7 system to study gene regulation and expression patterns. By placing a gene of interest under the control of the T7 promoter, they can precisely control its expression and observe its effects.

In Vitro Transcription: The T7 RNA polymerase can be used in vitro (outside a living organism) to synthesize RNA molecules from DNA templates containing the T7 promoter. This is widely used in generating RNA probes for various molecular biology techniques.

Gene Therapy: While still under development, there's potential for utilizing the T7 promoter in targeted gene therapy approaches, although challenges related to delivery and specificity need to be addressed.


4. Variations and Considerations



While "taatacgactcactataggg" strongly resembles the T7 promoter, minor variations may exist. Different variations might impact the efficiency of transcription. Optimizing the sequence to ensure maximum transcription efficiency for a specific application is a common practice in molecular biology labs. Factors such as the surrounding DNA sequence and the bacterial strain used also influence expression levels.

Furthermore, the choice of using the T7 promoter involves trade-offs. Its high strength can sometimes lead to the production of toxic proteins that harm the host bacteria. Careful experimental design and optimization are vital to mitigate these challenges.


5. Conclusion



The seemingly simple sequence "taatacgactcactataggg" represents a pivotal element in modern biotechnology, primarily due to its strong resemblance to the highly efficient T7 promoter. Its wide applicability in various areas, from protein production to gene expression studies, showcases its significant impact on scientific research and technological advancements. Understanding this sequence and its function is crucial for grasping fundamental aspects of molecular biology and its diverse applications.


FAQs:



1. What is the difference between a promoter and a gene? A promoter is a DNA region that initiates transcription of a gene. The gene itself contains the genetic code for a protein or RNA molecule. The promoter is essentially the "on" switch for the gene.

2. Why is the T7 promoter considered "strong"? The T7 promoter is considered strong because the T7 RNA polymerase binds to it with high affinity, leading to very efficient transcription.

3. Can the T7 promoter be used in organisms other than bacteria? While primarily used in bacteria, modified versions and adapted strategies are being explored for use in other organisms, though challenges remain.

4. Are there any disadvantages to using the T7 promoter? The high efficiency can lead to overwhelming protein production, potentially harming the host organism. Careful optimization is essential.

5. Where can I find more information on the T7 promoter and its applications? A literature search using keywords like "T7 promoter," "T7 RNA polymerase," and "recombinant protein expression" will yield numerous research articles and reviews providing detailed information.

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5' Primer Tags: A cheap way to fluorescently tag PCR products 5 Dec 2013 · Using "automated" DNA sequencers to score microsatellites requires that a fluorescent molecule (i.e., a "tag") be added. The most straightforward way to do this is to add a fluorescent molecule to the 5' end of one of the primers used in …

100002663PPS Primers Adapters - Thermo Fisher Scientific Stable for six months when stored at –20°C. Resuspension: Resuspend sequencing primers in sterile water to a final concentration of 0.1 μg/μl. Store resuspended primers at –20°C. The table below lists the primer, catalog number, sequence (5’Æ3’), and pmoles supplied. For technical support, email [email protected].

FLUORESCENT LABELED PRIMER SETS - MilliporeSigma Sequencing with fluorescent-labeled primers provides more even signal intensities among bases than dye terminator chemistries and generally produces read lengths of 450-500 bp with an accuracy of 98% or greater. Each set contains the specified primer labeled individually on the 5' end with one of the following dyes: FAM, JOE, ROX and TAMRA.

T7, T3 & SP6 Sequencing Primers - Gene Link Gene Link stocks various oligo dT primers, oligo dTVN primer, Oligo dT T7 primer, random primers, including an array of fluorescent dye labeled primers for genetic analysis using florescent detecting instruments. The C-12 amino labeled primers are ready to be conjugated to the investigator’s choice of NHS-activated ligand.

Mouse A4GALt ORF mammalian expression plasmid, N-His tag pCMV3-SP-N-His is recommended for constructing the N-His tag secretary and membrane proteins expression vector which containing a naïve signal peptide. An universal signal peptide is used to instead the naïve signal peptide.

Comparative performance of ELISA and dot blot assay for … Results: For dot blot assay, the minimum concentration to detect TRAb requir-ing 100 ng of antigen with antiserum diluted at 1:60. For diagnosing GD, the ELISA yielded a higher AUC compared with the dot blot assay (0.95 and 0.85, respectively).

Feasibility of Systemically Applied dsRNAs for Pest-Specific RNAi ... Gel electrophoresis was used to visualize the presence of exogenous dsRNA in treated seedling material and Sanger sequencing was used to further verify recovery of treatment dsRNAs.

pCMV3-N-FLAG Negative Control Vector (N-terminal FLAG-tagged) Negative control for the pCMV3-N-FLAG clone. Vector sequence is the same as pCMV3-N-FLAG, but multiple cloning sites are removed. Designed for mammalian expression, stable or transient. Hygromycin resistance gene for selection of stable cell lines. Each tube contains approximately 10μg of lyophilized plasmid.

PCR cloning with pASK-IBA, pPR-IBA and pEXPR-IBA vectors To avoid the fusion of such polylinker derived amino acids pASK-IBA, pPR-IBA and pEXPR-IBA vectors offer a general cloning strategy via Type IIS restriction enzymes, BsaI or Eco31I (NEB, MBI Fermentas).

Primer Design Considerations for Incorporating a T7 Promoter … Primer Design Considerations for Incorporating a T7 Promoter into a PCR Product for Subsequent in vitro Transcription/Translation. T7 promoter sequence (5′-TAA TAC GAC TCA CTA TAG GG-3′). Required for transcription of the DNA template. • ATG start codon (5′-ATG-3′) if not present in the sequence being amplified. Needed for translation initiation.

DIG Northern Starter Kit - MilliporeSigma T7 RNA Polymerase 5′TAATACGACTCACTATAGGG/X-mer or 5′TAATACGACTCACTATAGGA/X-mer T3 RNA Polymerase 5′AATTAACCCTCACTAAAGGG/X-mer Factors influencing stringency in hybridization • The degree of homology of the probe to the target RNA is an important factor for determining the appropriate hybridization conditions.

Maximizing transcription of nucleic acids with efficient T7 promoters Here, we used rapid ampli cation of cDNA 5′ends coupled fi with deep sequencing (5′ RACE-Seq) to test if sequences beyond the 3 nucleotide affect the activity of the T7 promoter.

pCR T7 TOPO TA Expression Kits - Thermo Fisher Scientific The pCR®T7 TOPO® TA Expression Kits are shipped on dry ice. Each kit contains three boxes as described below. Upon receipt, store the boxes as detailed below. This manual is supplied with the following kits. pCR®T7 TOPO TA Cloning® reagents (Box 1) are listed below. Note that the user must supply Taq polymerase. Store Box 1 at -20°C.

Champion pET302/NT-His and pET303/CT-His Vectors - Thermo … ChampionTM pET302/NT-His (5.7 kb) and pET303/CT-His (5.3 kb) vectors allow you to clone your gene of interest using restriction enzyme digestion and ligation.

In vitro Transcription T7 Kit (for siRNA Synthesis) PCR扩增产物作为模板 T7 启动子(TAATACGACTCACTATAGGG) 添加在sense primer 的5 ’端(这时,在T7启动子序列的上游添加6~10 个核苷酸可以更有效地促进T7 RNA polymerase 结合并合成RNA)、或者插入目的片段的质粒DNA 的T7 启动子上游设定sense primer 及anti-sense primer 进行PCR扩增。 扩增产物可使用NucleoSpin Gel and PCR Clean-up(Code No. 740609 …

T7 promoter sequencing primer, 20-mer - Fisher Sci The primer can be used to sequence DNA fragments located downstream from the specific T7 RNA polymerase promoter sequence. The primer may be unsuitable for sequencing certain plasmids that contain altered, although functionally active promoters.

Cloning and Mapping of Members of the MYM Family Tandem repeats of a novel, putative, zinc-binding motif (MYM) have been described within the products of two, highly homologous genes: ZNF198/RAMP/FIM and ZNF261/DXS6673E.

New Standard Primers - Eurofins Genomics T7 TAATACGACTCACTATAGGG. T7-981079 and T7minus1 T7term. CTAGTTATTGCTCAGCGGT pET-RP (CTAGTTATTGCTCAGCGG) T7-pET-mod CCCGCGAAATTAATACGACTCAC. Topo-1 TCGGATCCACTAGTAACG. U6_fwd GAGGGCCTATTTCCCATGATTCC. v5epitoperev CGTAGAATCGAGACCGAGGAGAGG. …

DNA Template Preparation for in vitro Transcription can be used as template for in vitro transcription. Linearized plasmid DNA, PCR products or cDNA can be used as templates for transcription if they contain a double-stranded RNA po. on of the promoter with respect to target sequence. The target sequence must be placed downstream of the promoter for sense RNA a.

Construction and characterization of an OmpH-deficient mutant of ... In the present paper, we report the construction of an attenuated P. multocida strain X- Table 1. Bacterial strains and plasmids used in this study.