Activity of GCK variants measured by functional complementation of an hxk1Δhxk2Δglk1Δ yeast strain.
Created
March 18, 2022
Last updated
June 14, 2022
Published June 14, 2022
Member of urn:mavedb:00000096
This study assessed the activity of human glucokinase (GCK) variants using functional complementation. The hxk1Δhxk2Δglk1Δ yeast strain shows reduced growth on glucose medium which is rescued by expression of human GCK. The growth of yeast cells expressing GCK variants on 0.2% glucose medium was used as a read-out of GCK activity.
The GCK sequence was divided into three regions that were mutagenized, assayed and sequenced in parallel. Each regional variant library was constructed by POPCode mutagenesis using pooled oligos each containing a central degenerate NNK codon. GCK variant activity was assayed using yeast complementation. Variants were sequenced using the TileSeq approach on an Illumina NextSeq550. The two replicates pre-selection were taken from the same pool of yeast transformants, while the two replicates post-selection were derived from the same pool of yeast transformants that were selected for GCK activity separately.
No associated publications.
Name: GCK
Type: Protein coding
Organism: Homo sapiens
Reference genome: hg38
Reference assembly: Other/Synthetic
Reference sequence: ATGTTAGATGACAGAGCAAGAATGGAAGCAGCCAAGAAGGAAAAGGTAGAACAAATATTAGCAGAATTTCAATTACAAGAAGAAGATTTGAAGAAAGTTATGAGAAGAATGCAAAAGGAAATGGATAGAGGTTTGAGATTAGAAACTCATGAAGAAGCTTCTGTTAAGATGTTGCCAACTTACGTTAGATCTACACCAGAAGGTTCAGAAGTTGGTGACTTTTTGTCATTAGATTTGGGTGGTACTAACTTCAGAGTTATGTTGGTTAAGGTTGGTGAAGGTGAAGAAGGTCAATGGTCTGTTAAGACAAAGCATCAAATGTACTCAATCCCAGAAGATGCTATGACTGGTACAGCAGAAATGTTGTTCGATTACATCTCTGAATGTATCTCAGATTTCTTGGATAAGCATCAAATGAAGCATAAGAAATTGCCATTGGGTTTTACTTTTTCTTTCCCAGTTAGACATGAAGATATCGATAAGGGTATCTTGTTGAACTGGACAAAGGGTTTTAAAGCTTCAGGTGCAGAAGGTAATAATGTTGTTGGTTTGTTGAGAGATGCTATTAAAAGAAGAGGTGACTTTGAAATGGATGTTGTTGCTATGGTTAACGATACTGTTGCAACAATGATCTCTTGTTACTACGAAGATCATCAATGTGAAGTTGGTATGATTGTTGGTACTGGTTGTAACGCATGTTACATGGAAGAAATGCAAAACGTTGAATTGGTTGAAGGTGACGAAGGTAGAATGTGTGTTAATACAGAATGGGGTGCTTTTGGTGACTCTGGTGAATTGGATGAATTCTTGTTGGAATACGATAGATTGGTTGATGAATCTTCAGCAAATCCAGGTCAACAATTGTACGAAAAGTTGATCGGTGGTAAATACATGGGTGAATTAGTTAGATTGGTTTTGTTGAGATTGGTTGATGAAAATTTGTTGTTCCATGGTGAAGCTTCTGAACAATTGAGAACTAGAGGTGCATTCGAAACAAGATTCGTTTCTCAAGTTGAATCAGATACTGGTGACAGAAAGCAAATCTATAACATCTTGTCTACATTAGGTTTAAGACCATCAACTACAGATTGTGATATTGTTAGAAGAGCTTGTGAATCTGTTTCAACTAGAGCTGCACATATGTGTTCTGCTGGTTTGGCAGGTGTTATTAATAGAATGAGAGAATCTAGATCAGAAGATGTTATGAGAATCACAGTTGGTGTTGATGGTTCTGTTTACAAGTTGCATCCATCTTTTAAAGAAAGATTCCATGCATCTGTTAGAAGATTGACTCCATCATGTGAAATCACTTTTATTGAATCTGAAGAAGGTTCAGGTAGAGGTGCCGCTTTAGTCAGTGCTGTTGCTTGTAAGAAGGCTTGTATGTTGGGTCAATAG
DOI: No associated DOIs
Raw reads:urn:mavedb:00000096-a-1
GCK activity measured by complementation
Activity of GCK variants measured by functional complementation of an hxk1Δhxk2Δglk1Δ yeast strain.