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Splicing reporters with 3′SS REPA or human G-tract mutant were created by replacing the corresponding upstream 3′SS of the middle constitutive exon of DUP175 as carried out previously [ 49].
The wild-type minigene L1-GHRexon8-L2 and the corresponding mutant were created to study the nucleotide change IVS7-6T>A, whish is located in the polypyrimidine tract before exon 8 and tested with the in vitro splicing assay.
Heterozygote mice expressing the I130T mutant were created as described by Fishman and colleagues [ 25] and were bred on a mixed background of CD1 and C57BL/6 (Gja1 tmicei) mice.
Homozygous diploid strains of the WT and UBP7 mutant were created by mating type switching using the YCp50::HO plasmid [ 92] and mating haploid strains of opposite mating type.
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In this study, a human TNF-α mutant was created and tested for its anti-tumor effects.
For this purpose, a genetically-engineered myoglobin (Mb) mutant was created by incorporating redox-active 3-amino-l-tyrosine (NH2Tyr) into its active site, replacing the distal histidine (H64) with NH2Tyr.
CΔ19 deletion mutant was created to remove a predicted nucleic acid binding domain.
An AXXLL containing Vpr mutant was created in the background of the p98CN009.8 Vpr.
The ΔhtsAΔisdE mutant was created for this study using allelic replacement procedures described by Bae and Schneewind [9].
To study the localization of PilP in PilQ-containing complexes within the membrane, a pilP deletion mutant was created.
Once a mutation occurs, a new mutant is created with fitness drawn randomly from the L possible neighbourhood values.
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