Construction of a Bacterial Artificial Chromosome Library for a Myxobacterium of the GenusCystobacterand Characterization of an Antibiotic Biosynthetic Gene Cluster

2005 ◽  
Vol 69 (7) ◽  
pp. 1372-1380 ◽  
Author(s):  
Zhiyang FENG ◽  
Jianhua QI ◽  
Takashi TSUGE ◽  
Yuichi OBA ◽  
Tetsuo KOBAYASHI ◽  
...  
Keyword(s):  
Bacterial Artificial Chromosome ◽  
Gene Cluster ◽  
Bacterial Artificial Chromosome Library ◽  
Artificial Chromosome ◽  
Biosynthetic Gene Cluster ◽  
Biosynthetic Gene

scholarly journals Simple cloning of large natural product biosynthetic gene cluster by CRISPR/Cas12a-mediated fast direct capturing strategy

2020 ◽  
Author(s):  
Mindong Liang ◽  
Leshi Liu ◽  
Weishan Wang ◽  
Xiaoqian Zeng ◽  
Jiakun Liu ◽  
...  
Keyword(s):  
Drug Discovery ◽  
Bacterial Artificial Chromosome ◽  
Gene Cluster ◽  
Bac Library ◽  
Gc Content ◽  
Artificial Chromosome ◽  
Biosynthetic Gene Cluster ◽  
Dna Fragments ◽  
Biosynthetic Gene ◽  
Direct Cloning

ABSTRACTDirectly cloning of biosynthetic gene clusters (BGCs) from even unculturable microbial genomes revolutionized nature products-based drug discovery. However, it is still very challenging to efficiently cloning, for example, the large (e.g. > 80kb) BGCs, especially for samples with high GC content in Streptomyces. In this study, by combining the advantages of CRISPR/Cas12a cleavage and bacterial artificial chromosome (BAC) library construction, we developed a simple, fast yet efficient in vitro platform for direct cloning of large BGCs based on CRISPR/Cas12a, named CAT-FISHING (CRISPR/Cas12a-mediated fast direct biosynthetic gene cluster cloning). It was demonstrated by the efficient direct cloning of large DNA fragments from bacterial artificial chromosomes or high GC (>70%) Streptomyces genomic DNA. Moreover, surugamides, encoded by a captured 87-kb gene cluster, was expressed and identified in a cluster-free Streptomyces chassis. These results indicate that CAT-FISHING is now poised to revolutionize bioactive small molecules (BSMs) drug discovery and lead a renaissance of interest in microorganisms as a source of BSMs for drug development.TABLE OF CONTENTSSIGNIFICANCE STATEMENTNatural products (NPs) are one of the most important resources for drug leads. One bottleneck of NPs-based drug discovery is the inefficient cloning approach for BGCs. To address it, we established a simple, fast and efficient BGC directed cloning method CAT-FISHING by combining the advantages of CRISPR/Cas12a (e.g. high specificity) and bacterial artificial chromosome (BAC) library (e.g. large DNA fragment and high GC content). As demonstrations, a series of DNA fragments ranging from 49 kb to 139 kb were successfully cloned. After further optimization, our method was able to efficiently clone and express an 87-kb long, GC-rich (76%) surugamides BGC in a Streptomyces chassis with reduced time-cost. CAT-FISHING presented in this study would much facilitate the process of NPs discovery.

Characterization of algG encoding C5-epimerase in the alginate biosynthetic gene cluster of Pseudomonas fluorescens

Gene ◽  
2001 ◽  
Vol 278 (1-2) ◽  
pp. 107-114 ◽  
Author(s):  
Antonella Morea ◽  
Kalai Mathee ◽  
Michael J. Franklin ◽  
Alessio Giacomini ◽  
Michael O'Regan ◽  
...  
Keyword(s):  
Pseudomonas Fluorescens ◽  
Gene Cluster ◽  
Biosynthetic Gene Cluster ◽  
Biosynthetic Gene

Cloning and characterization of the histidine biosynthetic gene cluster of Streptomyces coelicolor A3(2)

Gene ◽  
1990 ◽  
Vol 90 (1) ◽  
pp. 31-41 ◽  
Author(s):  
Danila Limauro ◽  
Alessandra Avitabile ◽  
Carmela Cappellano ◽  
Anna Maria Puglia ◽  
Carmelo B. Bruni
Keyword(s):  
Gene Cluster ◽  
Streptomyces Coelicolor ◽  
Biosynthetic Gene Cluster ◽  
Biosynthetic Gene

Molecular cloning and characterization of an ML-236B (compactin) biosynthetic gene cluster in Penicillium citrinum

2002 ◽  
Vol 267 (5) ◽  
pp. 636-646 ◽  
Author(s):  
Y. Abe ◽  
T. Suzuki ◽  
C. Ono ◽  
K. Iwamoto ◽  
M. Hosobuchi ◽  
...  
Keyword(s):  
Molecular Cloning ◽  
Gene Cluster ◽  
Biosynthetic Gene Cluster ◽  
Penicillium Citrinum ◽  
Biosynthetic Gene

Genetic localization and in vivo characterization of a Monascus azaphilone pigment biosynthetic gene cluster

2013 ◽  
Vol 97 (14) ◽  
pp. 6337-6345 ◽  
Author(s):  
Bijinu Balakrishnan ◽  
Suman Karki ◽  
Shih-Hau Chiu ◽  
Hyun-Ju Kim ◽  
Jae-Won Suh ◽  
...  
Keyword(s):  
Gene Cluster ◽  
Biosynthetic Gene Cluster ◽  
Biosynthetic Gene ◽  
Genetic Localization ◽  
In Vivo Characterization ◽  
Azaphilone Pigment
Sign in / Sign up

Export Citation Format

Share Document