DelsGate, a robust and rapid gene deletion construction method (Record no. 46472)
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| fixed length control field | 02489nam a2200289Ia 4500 |
| 003 - CONTROL NUMBER IDENTIFIER | |
| control field | MX-MdCICY |
| 005 - DATE AND TIME OF LATEST TRANSACTION | |
| control field | 20250625140706.0 |
| 040 ## - CATALOGING SOURCE | |
| Transcribing agency | CICY |
| 090 ## - LOCALLY ASSIGNED LC-TYPE CALL NUMBER (OCLC); LOCAL CALL NUMBER (RLIN) | |
| Classification number (OCLC) (R) ; Classification number, CALL (RLIN) (NR) | B-12261 |
| 008 - FIXED-LENGTH DATA ELEMENTS--GENERAL INFORMATION | |
| fixed length control field | 250602s9999 xx |||||s2 |||| ||und|d |
| 245 10 - TITLE STATEMENT | |
| Title | DelsGate, a robust and rapid gene deletion construction method |
| 490 0# - SERIES STATEMENT | |
| Volume/sequential designation | Fungal Genetics and Biology, 45(4), p.379-388, 2008 |
| 520 3# - SUMMARY, ETC. | |
| Summary, etc. | With the increasing availability of fungal genome sequences there is great demand for fast, simple high-throughput methods to generate constructs for gene deletion. Here we describe a method that combines PCR and Gateway cloning technology together with use of the I-SceI homing endonuclease to generate precise deletion constructs in a very simple, universal and robust manner in just 2 days. These constructs are then used to produce deletion mutants in the organism of interest following applicable methods for that species. In establishing this protocol we determined empirically that 1 kb was a suitable flank length to facilitate homologous recombination in our species of interest, Ustilago maydis. The method, which we have named DelsGate (Deletions via Gateway), consists of standard PCR of only the 50 and 30 1 kb gene flanks directly followed by in vitro Gateway cloning and final generation of the circular deletion construct by in vivo recombination in Escherichia coli. For use in DelsGate we have modified a Gateway cloning vector to include selectable markers for transformation of Ascomycetes and the Basidiomycete fungus U. maydis which causes corn smut disease. We have tested the reproducibility of the DelsGate approach by generating deletion constructs for 12 U. maydis genes. Although not tested here, the PCR and transformation steps of DelsGate should be well suited for high-throughput approaches to gene deletion construction in fungal species. DelsGate has the potential to be universal for all organisms with efficient transformation and homologous recombination systems. |
| 650 14 - SUBJECT ADDED ENTRY--TOPICAL TERM | |
| Topical term or geographic name entry element | GATEWAY |
| 650 14 - SUBJECT ADDED ENTRY--TOPICAL TERM | |
| Topical term or geographic name entry element | FUNGAL TRANSFORMATION |
| 650 14 - SUBJECT ADDED ENTRY--TOPICAL TERM | |
| Topical term or geographic name entry element | GENE DELETION |
| 650 14 - SUBJECT ADDED ENTRY--TOPICAL TERM | |
| Topical term or geographic name entry element | DELSGATE |
| 650 14 - SUBJECT ADDED ENTRY--TOPICAL TERM | |
| Topical term or geographic name entry element | USTILAGO MAYDIS |
| 700 12 - ADDED ENTRY--PERSONAL NAME | |
| Personal name | Garcáa-Pedrajas, M.D. |
| 700 12 - ADDED ENTRY--PERSONAL NAME | |
| Personal name | Nadal, M. |
| 700 12 - ADDED ENTRY--PERSONAL NAME | |
| Personal name | Kapa, L.B. |
| 700 12 - ADDED ENTRY--PERSONAL NAME | |
| Personal name | Perlin, M.H. |
| 700 12 - ADDED ENTRY--PERSONAL NAME | |
| Personal name | Andrews, D.L. |
| 700 12 - ADDED ENTRY--PERSONAL NAME | |
| Personal name | Gold, S.E. |
| 856 40 - ELECTRONIC LOCATION AND ACCESS | |
| Uniform Resource Identifier | <a href="https://drive.google.com/file/d/15SjYMlO_ByTfiVwb4s29rA7Hb2bfxSXo/view?usp=drivesdk">https://drive.google.com/file/d/15SjYMlO_ByTfiVwb4s29rA7Hb2bfxSXo/view?usp=drivesdk</a> |
| Public note | Para ver el documento ingresa a Google con tu cuenta: @cicy.edu.mx |
| 942 ## - ADDED ENTRY ELEMENTS (KOHA) | |
| Source of classification or shelving scheme | Clasificación local |
| Koha item type | Documentos solicitados |
| Lost status | Source of classification or shelving scheme | Damaged status | Not for loan | Collection | Home library | Current library | Shelving location | Date acquired | Total checkouts | Full call number | Date last seen | Price effective from | Koha item type |
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| Clasificación local | Ref1 | CICY | CICY | Documento préstamo interbibliotecario | 25.06.2025 | B-12261 | 25.06.2025 | 25.06.2025 | Documentos solicitados |