|
| Status |
Public on Sep 01, 2018 |
| Title |
G12-CBS_HP strain induced with methanol for 24 h_rep3 |
| Sample type |
RNA |
| |
|
| Source name |
GS115 harboring recombinant methionine adenosyltransferase (MAT) gene DS56 and the weak promoter G12 to downregulate CYS4 gene, S-
|
| Organism |
Komagataella phaffii GS115 |
| Characteristics |
phenotype: SAM low-producing
|
| Growth protocol |
Pichia pastoris strains GS115/3.5K, GS115/DS56 and G12-CBS were grown in BSM medium (1.82% K2SO4, 0.093% CaSO4, 2.68% H3PO4 (85%), 1.49% MgSO4•7H2O, 0.413% KOH, 4% glycerol), induced with methanol for 72 h, and supplied with L-Met after methanol induction for 24 h to promote SAM synthesis.
Pichia pastoris strainG12-CBS was grown in BSM medium induced with methanol for 24 h without L-Met feeding.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using the RNAeasy Kit (Qiagen, Valencia, CA, USA). The RNA quality and concentration were measured by NanoDrop ND-1000 and standard denaturing agarose gel electrophoresis.
|
| Label |
Cy3
|
| Label protocol |
Labeling was performed by NimbleGen Systems Inc., Madison, WI USA, following their standard operating protocol. See www.nimblegen.com.
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| |
|
| Hybridization protocol |
Hybridization was performed by NimbleGen Systems Inc., Madison, WI, USA following their standard operating protocol. See www.nimblegen.com.
|
| Scan protocol |
Slides were scanned at 5 μm/pixel resolution using an Axon GenePix 4000B scanner (Molecular Devices Corporation) piloted by GenePix Pro 6.0 software (Axon).Scanned images (TIFF format) were then imported into NimbleScan software (version 2.5) for grid alignment and expression data analysis.
|
| Description |
This sample is of engineered strain G12-CBS, which was grown in BSM medium, induced with methanol for 24 h without L-Met feeding. It is the third of three engineered strain G12-CBS biological replicates used in this experiment, each from separate cultures.
|
| Data processing |
Scanned images were extracted and normalized using NimbleScan software (version 2.5) through quantile normalization and the Robust Multichip Average (RMA) algorithm. All gene level files were then imported into Agilent GeneSpring GX software (version 11.5.1) for subsequent normalization by the quantile method and further analysis, including hierarchical clustering. Genes that at least 2 out of 12 samples have values greater than or equal to lower cut-off: 100.0 (All Targets Value) were chosen for data analysis.
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| |
|
| Submission date |
May 08, 2018 |
| Last update date |
Sep 01, 2018 |
| Contact name |
Xiulin Qin |
| E-mail(s) |
[email protected]
|
| Organization name |
Guangxi university
|
| Department |
College of Life Science and Technology
|
| Street address |
100 Daxue Road
|
| City |
Nanning |
| State/province |
Guangxi |
| ZIP/Postal code |
530004 |
| Country |
China |
| |
|
| Platform ID |
GPL17988 |
| Series (1) |
| GSE114152 |
Transcriptomic analysis of engineered and improved Pichia pastoris for S-adenosyl-l-methionine (SAM) production |
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