A new method for simple and fast clone checking is described. We combined the Pyrosequencing technology with a preprogrammed nucleotide dispensation strategy for fast analysis of DNA constructs. To test this method, the N-terminus region of plasmids constructed for the production of recombinant apyrase was analyzed. Of the ten plasmids tested, seven constructs were correct, two constructs showed one base deletion, and one construct showed deletion of a 195 bp fragment. The preprogrammed nucleotide dispensation strategy allowed the identification of the sequence downstream of the deletions. Thus, this method determines both the location and nature of possible artifacts.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1002/elps.200305434 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Visual codon: a user-friendly Python program for viewing and optimizing gene GC content.
PeerJ
December 2024
Life Science College, Fujian Agriculture and Forestry University, Fuzhou, Fujian, China.
Due to the codon bias of different species, codon optimization is usually carried out in the process of heterologous protein expression. At present, there are a variety of codon optimization tools. However, the optimized sequences may still have high or low points of local guanine and cytosine (GC) content, which is not conducive to the primer design of gene subcloning, and also makes it difficult to perform the experiment of synthesizing the whole gene with DNA fragments by polymerase chain reaction (PCR) reaction.
View Article and Find Full Text PDFScreening and identification of viruses in termites: behavior-regulating roles of RcBsV and RcBV.
Pest Manag Sci
December 2024
Hubei Insect Resources Utilization and Sustainable Pest Management Key Laboratory, Huazhong Agricultural University, Wuhan, China.
Background: Termites live underground in a social setting having continuous contact with microorganism. However, there is no comparative study on virus diversity and relative abundance between termites, castes and body parts in termites. To address this gap, pseudergates of Cryptotermes declivis, workers of the Odontotermes formosanus, and workers, soldiers and alates of the Reticulitermes chinensis were used as experimental materials to perform virome sequencing, virus annotations and their relative abundance analysis.
View Article and Find Full Text PDFMetagenomics approaches in the discovery and development of new bioactive compound of 8-demethoxy-10-deoxysteffimycin from mangrove sediments.
3 Biotech
December 2024
Department of Cariology, Saveetha Dental College and Hospitals, Saveetha Institute of Medical and Technical Sciences, Saveetha University, Chennai, India.
A metagenomic library consisting of 15,000 clones was constructed from the mangrove sediment. An antimicrobially active clone from the metagenomic library PS49 was identified by function- based screening. This paper presents the results of the biochemical characterization and metagenomic library screening of the marine-derived antibiotic, 8-demethoxy-10-deoxysteffimycin.
View Article and Find Full Text PDFIdentification of a novel alternate promoter element in the pheST operon of Escherichia coli.
Mol Biol Rep
October 2024
Department of Molecular Biology, School of Biological Sciences, Madurai Kamaraj University, Madurai, 625 021, India.
Background: Earlier work in this laboratory revealed that fitA was same as pheS as a recombinant clone, pSRJ5R1 harboring pheS gene complemented transcription-defective fitA76 Ts (temperature sensitive) mutant. However, this clone lacked the native promoter (NP) of pheST operon. A putative - 10 promoter like element was suggested to act as promoter in this clone.
View Article and Find Full Text PDFDevelopment and validation of a multi-marker liquid bead array assay for the simultaneous detection of and hotspot mutations in single circulating tumor cells (CTCs).
Heliyon
October 2024
Analysis of Circulating Tumor Cells Lab, Laboratory of Analytical Chemistry, Department of Chemistry, National and Kapodistrian University of Athens, 15771 Athens, Greece.
Article Synopsis
- The study presents a new liquid bead array assay that detects specific mutations in circulating tumor cells (CTCs) from metastatic breast cancer (MBC) patients, which can help identify tumor progression and therapy resistance.
- The assay utilizes advanced techniques like enzymatic mutation enrichment and multiplex PCR, allowing for highly sensitive (0.1% mutation detection limit) and specific identification of mutations in single CTCs.
- Validation of the assay showed that it can analyze 96 single cells at once, making it both efficient and sample-conserving, thereby providing crucial insights into the genetic landscape of tumors.
Want AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!