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Our lab has shown that single-stranded oligonucleotides (ODN) can be used to repair genes with single base mutations within the context of the native chromosome.
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The number of sequences in the distinct homologous families ranged from 6 110 (Table 1) with few base mutations within a family.
By itself, such a mechanism is incapable of explaining similarities between complementary-base mutations within each of these two classes.
To determine if the GLS is required for normal gurken gene function, we created a rescue construct, called grkGLSmut, that is identical to grkwt, except for the inclusion of 12 single-base mutations, all within the GLS.
Three base-pair mutations within the NaCDEI (21 23a and 24 26c) and NaCDEII (45 47t) of NcCEN1 showed substantial decreases in yeast colony formation and high copy numbers of minichromosomes (>70 per cell), indicative of inefficient segregation.
The PG-1 allele is the result of a spontaneous single-base mutation within a AtMYB90 transgene that acts as a dominant-negative 'repressor' of pigment production in tobacco flowers.
In addition, a second construct was created with a 5 base pair mutation within the target seed site (Psi/miR34amut).
Leukocyte adhesion deficiency type 1 (LAD-1) is an autosomal recessive disorder based on point mutations within the β2 integrin, resulting in either its absence on the cell surface or the inability to associate with the corresponding α subunit.
A single-base nonsense mutation within the coding region of an active Arabidopsis AtMYB90 transgene (the PG-1 allele) was found to convert the R2R3-myb gene from a transcriptional activator of plant-wide anthocyanin biosynthesis to a dominant-negative allele that was able to interfere with normal tobacco pigment production within flower petals.
After quality control, sequence error rate was estimated as the percentage of single-base mutations in the rpl32- trnL region within individuals and was found to be approximately 0.13% (see Methods).
We have developed a novel concept for immunogen construction based on introduction of massive mutations within the epitopes targeting antigenically variable pathogens and diseases.
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