A cloning method to identify caspases and their regulators in yeast: Identification of Drosophila IAP1 as an inhibitor of the Drosophila caspase DCP-1

The peptides from the ranacyclin family share similar active disulphide loop with plant-derived Bowman–Birk type inhibitors, some of which have the dual activities of trypsin inhibition and antimicrobial. Herein, a novel Bowman–Birk type trypsin inhibitor of the ranacyclin family was identified from the skin secretion of broad-folded frog (Sylvirana latouchii) by molecular cloning method and named as SL-BBI. After chemical synthesis, it was proved to be a potent inhibitor of trypsin with a Ki value of 230.5 nM and showed weak antimicrobial activity against tested microorganisms. Modified analogue K-SL maintains the original inhibitory activity with a Ki value of 77.27 nM while enhancing the antimicrobial activity. After the substitution of active P1 site to phenylalanine and P2′ site to isoleucine, F-SL regenerated its inhibitory activity on chymotrypsin with a Ki value of 309.3 nM and exhibited antiproliferative effects on PC-3, MCF-7 and a series of non-small cell lung cancer cell lines without cell membrane damage. The affinity of F-SL for the β subunits in the yeast 20S proteasome showed by molecular docking simulations enriched the understanding of the possible action mode of Bowman–Birk type inhibitors. Further mechanistic studies have shown that F-SL can activate caspase 3/7 in H157 cells and induce apoptosis, which means it has the potential to become an anticancer agent.

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A new cloning method for antibody-forming lymphoblastoid cells

Journal of Immunological Methods ◽

10.1016/0022-1759(86)90234-6 ◽

1986 ◽

Vol 94 (1-2) ◽

pp. 201-208 ◽

Cited By ~ 7

Author(s):

Slawomir Kosinski ◽

Ulrich Hämmerling

Keyword(s):

Lymphoblastoid Cells ◽

Cloning Method

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IRDL Cloning: A One-Tube, Zero-Background, Easy-to-Use, Directional Cloning Method Improves Throughput in Recombinant DNA Preparation

PLoS ONE ◽

10.1371/journal.pone.0107907 ◽

2014 ◽

Vol 9 (9) ◽

pp. e107907 ◽

Cited By ~ 4

Author(s):

Jiancai Wang ◽

Ronghua Xu ◽

Aizhong Liu

Keyword(s):

Recombinant Dna ◽

Dna Preparation ◽

Cloning Method ◽

Directional Cloning

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Piglets born from handmade cloning, an innovative cloning method without micromanipulation

Theriogenology ◽

10.1016/j.theriogenology.2007.07.021 ◽

2007 ◽

Vol 68 (8) ◽

pp. 1104-1110 ◽

Cited By ~ 75

Author(s):

Y. Du ◽

P.M. Kragh ◽

Y. Zhang ◽

J. Li ◽

M. Schmidt ◽

...

Keyword(s):

Cloning Method ◽

Handmade Cloning

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Yeast identification

Nature ◽

10.1038/283799b0 ◽

1980 ◽

Vol 283 (5749) ◽

pp. 799-800

Author(s):

B.H. Kirsop

Keyword(s):

Yeast Identification

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The alpha-mating type locus of Cryptococcus neoformans contains a peptide pheromone gene

Molecular and Cellular Biology ◽

10.1128/mcb.13.3.1962-1970.1993 ◽

1993 ◽

Vol 13 (3) ◽

pp. 1962-1970

Author(s):

T D Moore ◽

J C Edman

Keyword(s):

Cryptococcus Neoformans ◽

Mating Type ◽

Cell Types ◽

Yeast Cells ◽

Mating Types ◽

Type Locus ◽

Reading Frame ◽

Cloning Method ◽

Opportunistic Fungal Pathogen ◽

Specific Fragment

The opportunistic fungal pathogen Cryptococcus neoformans has two mating types, MATa and MAT alpha. The MAT alpha strains are more virulent. Mating of opposite mating type haploid yeast cells results in the production of a filamentous hyphal phase. The MAT alpha locus has been isolated in this study in order to identify the genetic differences between mating types and their contribution to virulence. A 138-bp fragment of MAT alpha-specific DNA which cosegregates with alpha-mating type was isolated by using a difference cloning method. Overlapping phage and cosmid clones spanning the entire MAT alpha locus were isolated by using this MAT alpha-specific fragment as a probe. Mapping of these clones physically defined the MAT alpha locus to a 35- to 45-kb region which is present only in MAT alpha strains. Transformation studies with fragments of the MAT alpha locus identified a 2.1-kb XbaI-HindIII fragment that directs starvation-induced filament formation in MATa cells but not in MAT alpha cells. This 2.1-kb fragment contains a gene, MF alpha, with a small open reading frame encoding a pheromone precursor similar to the lipoprotein mating factors found in Saccharomyces cerevisiae, Ustilago maydis, and Schizosaccharomyces pombe. The ability of the MATa cells to express, process, and secrete the MAT alpha pheromone in response to starvation suggests similar mechanisms for these processes in both cell types. These results also suggest that the production of pheromone is under a type of nutritional control shared by the two cell types.

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Comparative Evaluation of the BD Phoenix Yeast ID Panel and Remel RapID Yeast Plus System for Yeast Identification

Canadian Journal of Infectious Diseases and Medical Microbiology ◽

10.1155/2016/4094932 ◽

2016 ◽

Vol 2016 ◽

pp. 1-4 ◽

Cited By ~ 2

Author(s):

Michelle L. Grant ◽

Shobha Parajuli ◽

Raquel Deleon-Gonsalves ◽

Raghava Potula ◽

Allan L. Truant

Keyword(s):

Comparative Evaluation ◽

Concordance Rate ◽

Becton Dickinson ◽

Yeast Identification

Becton Dickinson Phoenix Yeast ID Panel was compared to the Remel RapID Yeast Plus System using 150 recent clinical yeast isolates and the API 20C AUX system to resolve discrepant results. The concordance rate between the Yeast ID Panel and the RapID Yeast Plus System (without arbitration) was 93.3% with 97.3% (146/150) and 95.3% (143/150) of the isolates correctly identified by the Becton Dickinson Phoenix and the Remel RapID, respectively, with arbitration.

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