Navigation Links
Transformation of Filamentous Fungi by Microprojectile Bombardment

s for microprojectile bombardment were compared with literature data on Biolistic transformation of other fungal species.


Materials and Methods
Strain and plasmid
Aspergillus nidulans strain A773 (pyrG89, wA3, pyroA4) was obtained from the Fungal Genetics Stock Center, Dept. of Microbiology, University of Kansas Medical Center, Kansas City, Kansas, and maintained on solid YG medium [0.5% yeast extract, 2% glucose, 2% agar, pH 6.3] supplemented with 0.12% uracil and 0.12% uridine. Plates were incubated for 68 days at 37 C at which time conidia were harvested in 0.9% sterile saline. The resuspended conidia were sonicated in an ultra sound sonicator for 1 min prior to being used for protoplast formation or for microprojectile bombardment.

Plasmid pRG-1, a 5 kb plasmid containing the Neurospora crassa pyr4 gene cloned into pUC9 was used for transformation experiments. This plasmid has been shown to complement the pyrG89 mutation in A. nidulans (Ballance et al ., 1983; Waring et al ., 1989).

Biolistic transformation
Intact conidia were spread onto 10 ml of solid YG medium (in a 100 x 15 mm petri dish), briefly air-dried under sterile conditions, and used for microprojectile bombardment within 13 hours. Each plate was bombarded twice. Tungsten particles (M5, M10, or M17 particles, Bio-Rad Laboratories, Hercules, California) were prepared and coated with plasmid DNA as described by Daniell (1993). The following were added in order to a 1.5 ml microcentrifuge tube: 25 l tungsten particle suspension (1.5 mg in 50% glycerol), 5 l plasmid DNA (0.5 g/l), 25 l 2.5 M CaCl2, and 5 l 1 M spermidine free base. After eac
'"/>

Source:


Page: All 1 2 3 4 5 6 7 8 9

Related biology technology :

1. Highest Possible Transformation Efficiencies for High-Throughput Applications
2. New Competent Cells for Highest Transformation Efficiencies
3. Optimization of Biolistic Transformation Using the Helium-Driven PDS-1000/He System
4. Sub-Micron Gold Particles Are Superior to Larger Particles for Efficient Biolistic Transformation of Organelles and Some Cells
5. Transformation of Nematodes With the Helios Gene Gun
6. Transformation of Filamentous Fungi by High-Voltage Electroporation
7. Detection of Reporter Gene Activity in Cell Cultures and Murine Epidermis After Helios Gene Gun-Mediated Particle Bombardment
Post Your Comments:
(Date:4/30/2015)... Cytokinetics, Incorporated (Nasdaq: CYTK) reported total research ... $4.4 million, compared to $8.0 million during the same period ... was $8.9 million, or $0.23 per basic and diluted share. ... period in 2014, of $8.7 million, or $0.27 per basic ... cash equivalents and investments totaled $117.5 million. , "During ...
(Date:4/30/2015)... The following is an open letter sent April 30, 2015 ... (NYSE: CO ) from Jayhawk Capital. We ... China Cord Blood Corp (" China Cord ") will REJECT ... Golden Meditech Holdings Co Ltd ("Golden Med"). Certainly, a going ... Med and Mr. Kam, the Chairman of both China ...
(Date:4/30/2015)... 30, 2015  Asterias Biotherapeutics, Inc. (NYSE MKT: AST), ... regenerative medicine, today announced that it will release first ... 2015 after the close of the U.S. financial markets. ... on Thursday, May 7, 2015 at 4:30 p.m. Eastern ... recent corporate developments. For both "listen-only" participants ...
(Date:4/30/2015)... FRANCE (PRWEB) April 30, 2015 ... and at http://www.lespausa.com , Phytomer’s DOUCEUR ... sensitive and reactive skin while battling the weakening ... This restorative cream is formulated with star ingredient ... via biotechnology in a complex of soothing minerals ...
Breaking Biology Technology:Cytokinetics, Incorporated Reports First Quarter 2015 Financial Results 2Cytokinetics, Incorporated Reports First Quarter 2015 Financial Results 3Cytokinetics, Incorporated Reports First Quarter 2015 Financial Results 4Cytokinetics, Incorporated Reports First Quarter 2015 Financial Results 5Cytokinetics, Incorporated Reports First Quarter 2015 Financial Results 6Cytokinetics, Incorporated Reports First Quarter 2015 Financial Results 7Cytokinetics, Incorporated Reports First Quarter 2015 Financial Results 8April 30, 2015 Open Letter to the Board of China Cord Blood Corp from Jayhawk Capital 2April 30, 2015 Open Letter to the Board of China Cord Blood Corp from Jayhawk Capital 3April 30, 2015 Open Letter to the Board of China Cord Blood Corp from Jayhawk Capital 4April 30, 2015 Open Letter to the Board of China Cord Blood Corp from Jayhawk Capital 5Asterias Biotherapeutics to Report First Quarter Results on May 7, 2015 2Asterias Biotherapeutics to Report First Quarter Results on May 7, 2015 3Phytomer introduces DOUCEUR INTEMPORELLE Restorative Shield Cream 2Phytomer introduces DOUCEUR INTEMPORELLE Restorative Shield Cream 3
... 23 ArunA Biomedical, Inc., announced ... MN, giving Neuromics the right to non-exclusively market ... and Neural Culture Medium to support applications in ... an exclusive worldwide license to develop and commercialize ...
... 23 BioDuro and AstraZeneca announced today that ... discovery research for Respiratory and Inflammation indications. BioDuro ... of discovery chemistry, discovery biology, ADMET*, and DMPK** ... discovery and shorten development timelines. "After a successful ...
... Nereus Pharmaceuticals, Inc., a pioneer in drug discovery ... is conducting a randomized Phase 2 clinical trial ... combination with standard chemotherapy (docetaxel) in patients with ... follows on positive outcomes in the Phase 1 ...
Cached Biology Technology:ArunA Biomedical, Inc. Announces Alliance with Neuromics for Distribution of Normal Human Neural Cells 2BioDuro and AstraZeneca Expand Collaboration to Optimize Discovery Research for Respiratory and Inflammation Indications 2Nereus Pharmaceuticals Initiates ADVANCE, a Randomized Phase 2 Clinical Trial of NPI-2358 in Non-Small Cell Lung Cancer 2Nereus Pharmaceuticals Initiates ADVANCE, a Randomized Phase 2 Clinical Trial of NPI-2358 in Non-Small Cell Lung Cancer 3
(Date:4/2/2015)... -- According to preliminary estimates Fingerprint Cards, ... than 125 MSEK. This exceeds the guidance provided by ... for Q1 2015 would be in line with the ... The operating result for the first quarter is estimated ... will as previously communicated be published on May 21. ...
(Date:4/1/2015)... OXFORD, Conn. , Apr. 1, 2015  NXT-ID, ... a biometric authentication company focused on the growing mobile ... number of Wocket smart wallets is underway to early ... early access group includes usage at retail outlets including ... CVS Pharmacy. Users report Wocket was accepted at all ...
(Date:3/31/2015)... 2015  Elephant Talk Communications Corp. (NYSE MKT: ETAK) ... Software Defined Network Architecture (ET Software DNA® 2.0) platforms ... approximately $20.4 million for the year ended December 31, ... the year ended December 31, 2013. ... completed its multi-year transition away from its legacy landline ...
Breaking Biology News(10 mins):Fingerprint Cards Revises Revenues for Q1 2015 2NXT-ID Reports Second Shipment of Wocket Smart Wallets Underway to Early Access Pre-order Customers and Provides User Feedback 2NXT-ID Reports Second Shipment of Wocket Smart Wallets Underway to Early Access Pre-order Customers and Provides User Feedback 3NXT-ID Reports Second Shipment of Wocket Smart Wallets Underway to Early Access Pre-order Customers and Provides User Feedback 4Elephant Talk Reports 2014 Fourth Quarter and Year End Financial Results 2Elephant Talk Reports 2014 Fourth Quarter and Year End Financial Results 3Elephant Talk Reports 2014 Fourth Quarter and Year End Financial Results 4Elephant Talk Reports 2014 Fourth Quarter and Year End Financial Results 5Elephant Talk Reports 2014 Fourth Quarter and Year End Financial Results 6
... have uncovered how members of one family of antibiotics ... knowledge may help drug developers make slight changes to ... strains of bacteria, said Irina Artsimovitch, a study co-author ... University. , The antibiotics studied belong to the rifamycin ...
... Rice University, the Baylor College of Medicine, the University of ... have created a new class of magnetic resonance imaging (MRI) ... than the best in clinical use. , The new agents ... gadolinium, that is given to more than a quarter of ...
... About five to seven million years ago, when the ... similar to rutabagas and turnips may have been one ... Laden of the University of Minnesota and Richard Wrangham ... organs like roots and tubers must have sustained our ...
Cached Biology News:Researchers find how some antibiotics kill bacteria 2Researchers find how some antibiotics kill bacteria 3'Gadonanotubes' greatly outperform existing MRI contrast agents 2The roots of civilization trace back to ... roots 2
6B9...
Our original highly-purified, robust Taq Polymerase which delivers consistently high yields and minimal background...
... - 5 units/µL ,• High specific activity ... Leaves an 'A' overhang ,• Processes ... very high activity in primer extension and ... quality of the enzyme, each lot has ...
Aspergillus (IgG) Ab EIA Sample Size: 100 l...
Biology Products: