Week 2 HW: DNA Design

Part 1: Define your expression target

My repository is Kelly_Mycelium: I would like to use Mycelium, the vegetative network of fungi, to form a naturally porous

What molecule are you expressing?

I would choose Vmh2, a Class I hydrophobin, because it is specifically associated with the vegetative mycelium of P. ostreatus. UniProt reports that Vmh2 is involved in mycelial surface hydrophobicity and biofilm formation, is secreted/cell-wall associated, and self-assembles into rodlet structures

What is the sequence?

https://www.uniprot.org/uniprotkb/A0A067N4H9/entry#sequences

Native protein sequence: P. ostreatus Vmh2, UniProt A0A067N4H9

The DNA coding sequence will be synthesized as a host-compatible, codon-optimized sequence while encoding the intended Vmh2 protein sequence.

MFSRVMFCTFLILPLLAAATAIPRTDTPSCSTGSLQCCSSVQKASDPLVGIIVALLGIVLGPLDLNVGLTCSPITVIGVGGTSCTQQTVCCTGNNFDGLIVAGCSPINIGL

What host or chassis?

The initial expression chassis will be Saccharomyces cerevisiae. Yeast will serve as a model system for investigating production and extracellular behaviour of the fungal hydrophobin before considering translation of the findings to P. ostreatus. This separation allows the molecular component of the hypothesis to be investigated independently from the more complex fungal-material system.

Where does the product need to go?

The target localization is extracellular/secretion-associated. Native Vmh2 is annotated as secreted and cell-wall associated in P. ostreatus. Therefore, the expression design should support secretion rather than simple intracellular accumulation. Extracellular localization matters because the proposed mechanism involves hydrophobin interaction with interfaces and fungal surface organization.

Part 2: Design your genetic parts

Yeast promoter → 5′ UTR/Kozak → Vmh2 CDS → secretion signal → terminator/poly(A) → yeast-compatible backbone

Part 3: Assemble your construct

The design needs more work.