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Promoter Library

Introduction

As a part of our project we wish to create a library of SigmaA promoters and characterise them based on their stregnths.

Novelty in this sub-project

A library of promoters with different strengths would be very useful for engineering biological devices. Promoters' strength can be compared with their PoPS values. However measuring the PoPS is not a trivial process. We can instead meausure their strengths relatively.

Design

Template used to create the promoter sequences: Prefix...UpSpacer...-35...PromSpacer...-10...DownSpacer..Suffix

• BBPrefix - StandardBioBrickPrefix EcoR1 and Xba1 - gaattcgcggccgcttctagag
• UpSpacer - attta - consensus sigA 5 bases 5'
• -35 - TTGACA
• PromSpacer - length 17 - consensus ttttatttaaattatga
• -10 - TATAAT
• DownSpacer (from ackA)- ggaaaag
• BBSuffix - Standard BioBrick suffix Spe1 and Pst1 - tactagtagcggccgctgcag

Variances

We designed three variances to create library of promoters with different strengths.

1. Degenerating -35 and -10 sequences.

 BBPrefix...UpSpacer...N-35... PromSpacer...N-10...DownSpacer..BBSuffix

1. BBPrefix...UpSpacerNN...-35...NNPromSpacerNN...-10...NNDownSpacer..BBSuffix
2. BBPrefix...UpSpacer...-35... 18N’s...-10...DownSpacer..BBSuffix

Variance 1

In this variance we only degenerate -35 and -10 sequences.

Variance 2

Variance 3

Lab Work Strategies

Other Presentations and Diagrams