Endy:Mycoengineering Subgroup

Week of 7/24/17

"Rolando"

• Build pSB3K5_sfGFP_AgroT and transform into Agro., image cells
• Run G. lucidum ATMT
• Retreat poster
• Quals packet
• Test PLL treated cover slip fungi cell immobilization
• Test nanopipette injection of immobilized fungi cells

Week of 7/24/17

"Rolando"

• build vector with sfGFP driven by 5' UTR of virE. (should report when I am sufficiently inducing the A. tum. Type IV secretion system).
• Prepare for meeting with X.

last week:

• advised REUs.
• identified suitable standard vector (sfGFP driven by J23119) to transform into A. tum.
• designed and ordered primers/REs for building pHg_GL_EXP backbone, pHg_GL_virEsfGFP.

Week of 7/17/17

"Rolando"

Build "vir" induction reporter vector.

• tansforma agro.
• build vector with gfp into CAMBIA based vector

last week:

spent my time advising/mentoring REUs

Week of 7/10/2017

Rolando

Build vir induction reporter vector.

• Transform Agro. with E. coli GFP/RFP reporter vector.
• Build vector with GFP driven by vir gene promoter.

Run electroporation experiment (w/Isaac).

• Electroporate pHg_LB, pHg_GL and pCbx.

Protoplasts (w/Isaac).

• Design protoplast optimization (w/close attention to regeneration).
• Run experiment.

Isaac

Last week:

Sequence Verifying pHg and pCbx mycobricks

Protoplasting and plating G. lucidum

Starting liquid and plated cultures

Prototyping a climate recording grow box

This week:

Protoplasting G. lucidum for transformation and troubleshooting

Electroporation of G. Lucidum for transformation

Agrobacterium mediated transformation of G. lucidum

Design of pHg_GFP and pHg_Cas9 plasmids