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Analysis of protein interactions with Tlpsig2.3.4 using the yeast two-hybrid system.

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posted on 2014-01-02, 02:43 authored by Hossinur Rahman, Rebecca M. King, Lucy K. Shewell, Evgeny A. Semchenko, Lauren E. Hartley-Tassell, Jennifer C. Wilson, Christopher J. Day, Victoria Korolik

Testing for interactions of Tlpsig2.3.4 fused to the GAL-4 activation domain (AD- Tlpsig2.3.4) and GAL-4 binding domain (BD- Tlpsig2.3.4) with all chemotaxis proteins and individual domains fused to the GAL-4 DNA binding domain and GAL-4 activation domain respectively. Co-transformation of AD-CheW and BD-CheA was used as a positive control when testing for interactions, with a result of +++.

+++: cream, dense growth of >75% of co-transformants observed on high stringency media and intermediate stringency media.

++: cream − pink moderate growth for ∼75% or more of co-transformants observed on high stringency media and intermediate stringency media.

+: cream − pink light growth for ∼50% or more of co-transformants observed on high stringency media and/or intermediate stringency media.

0: no growth of co-transformants observed.

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