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From 2010.igem.org

Contents 1 Hassan 1.1 Folate 1.2 Possible Genes 2 Mimmi 2.1 MITF 2.1.1 - Gel 2.2 MITF + yCCS and RFP 2.2.1 Verification PCR 3 Andreas

Hassan

Folate

below links and articles are meant to be study when I'm finished with interaction map and prove of concept!

• [http://www.ncbi.nlm.nih.gov/pubmed/12761857]
• [http://www.ncbi.nlm.nih.gov/pubmed/17307853,18403248,17698004,17220358,16359333,10201405,15489424,2254281,15865426,17923103,11041843,2185835,11371182,9287033,1958664,8208138,8019142,8408018,8634297,1098865?report=docsum]
• [http://www.ncbi.nlm.nih.gov/pubmed/15611104]
• [http://www.ncbi.nlm.nih.gov/pubmed/17698004]
• [http://www.ncbi.nlm.nih.gov/pubmed/8208138]
• [http://www.genome.jp/kegg-bin/show_pathway?org_name=eco&mapno=00790&mapscale=&show_description=hide]

Possible Genes

• decrease in CAT (Catalase) : [http://www.ncbi.nlm.nih.gov/pubmed/10537016][http://onlinelibrary.wiley.com/doi/10.1034/j.1600-0749.2003.00087.x/abstract]
• increase in SOD (SOD3) : [http://www.ncbi.nlm.nih.gov/pubmed/12692376]
• possible genes involved in Melanocytes adhesion: "An imbalance in cell adhesion molecules was also observed in vitiligo melanocytes. Differentially downregulated genes in vitiligo melanocytes included integrin alpha 7, integrin beta-like 1, integrin beta 7 and catenin alpha-like 1 (cadherin-associated protein). The adhesion between the melanocytes and the basal membrane is mediated by integrins and the contacts between melanocytes and keratinocytes are mediated by cadherins (Hara et al., 1994; Tang et al., 1994; Zambruno et al., 1993). The importance of regulated expression levels of cadherins have also been shown during migration of melanoblasts (Nishimura et al., 1999). It is difficult to predict if this specific downregulation of adhesion molecules impairs the attachment of the melanocytes in the basal layer of the epidermis; however, detachment and transepidermal loss of melanocytes have previously been reported in vitiligo vulgaris (Gauthier et al., 2003)."[http://www.ncbi.nlm.nih.gov/pubmed/18426409] May also look at [http://www.ncbi.nlm.nih.gov/pubmed/17850512] if more interested.

And those from July 22 with data from planning will bring enough explanation for interaction map.

I think by end of this week a wall of text will appear in modelling page!

Mimmi

MITF

- Gel

• No products!

--> Trying more cycles of low annealing temperature

MITF + yCCS and RFP

Verification PCR

Mix	(µl)	X5	X3		yCCS/RFP			MITF
sH2O	22.5	112.5	67.5		pSB1_VF2			MITF_F
F primer	1	5	3		pSB1_VR			MITF_R
R primer	1	5	3		970/925bp			~1300bp
DNA	0.5	4X0.5	2X0.5
tot	25				conditions			conditions
					time	°C		time	°C
					2m	94		2m	94
					30s	94	)	30s	94	)
					30s	60	> 30 cycles	30s	60	> 30 cycles
					2m	72	)	2m40s	72	)
					10m	72		10m	72
					oo	10		oo	10

Andreas