Re: [DIYbio] Re: query from PBS NewsHour

Synthetic forward genetics? Awesome! I heartily endorse such excellent and ambitious science! :)

Tom Randall <tarandall@gmail.com> wrote:

>
>
>
>> (1) Is anyone actually doing DIY work on synthetic biology? What
>makes it
>> DIY (e.g. taking place in a community lab)? What makes it synbio
>(and not
>> just biology or molecular biology)?
>>
>>
>> Thanks,
>> Jason
>>
>>
>> I have a lab at home and will likely soon be doing something that
>loosely
>might be considered synbio, but only because I intend on ordering a
>sequence from one of the companies like Blue Heron.
>Specifically the project is that I have a morphological mutant in my
>organism of interest, an ascomycete fungus Neurospora crassa. I am
>interested in determining the gene responsible for the phenotype of
>this
>mutation. It has been genetically mapped so I know within 20-40 kb
>where it
>is. I have in the past done Sanger sequencing on a candidate gene,
>which
>found no mutation in or around that particular gene so decided to
>simply
>get the whole genome sequenced. Hopefully then I can compare that
>sequence
>to reference and find a SNP/indel responsible. If that is possible, I
>intend to order a sequence of ~ 1 kb around this mutation, including
>the
>candidate mutant allele, and transform this into a strain with the wild
>
>type allele of this candidate gene in order to generate a replacement
>of
>the wt allele with the mutant allele by homologous recombination in
>order
>to re-create the original phenotype and thus confirm (or not) the
>genetic
>lesion that resulted in the original mutant allele. I have sent the DNA
>
>from this mutant strain to Operon, but only last week so the sequencing
>is
>still is in progress. I would say this is only loosely synbio because I
>am
>simply going to be using a synthetic construct as a convenient tool in
>the
>context of what is really a molecular genetics/bioinformatics project.
>
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