Difference between revisions of "Part:BBa K4765120"

Revision as of 15:07, 4 October 2023

ribozyme test: T7 leaky expression

Introduction

This composite part is utilized to assess the cleavage efficiency of Twister P1， stem-loop’s ability of preventing mRNA degradation and the strength of different ribosome binding site (RBS). It is regulated by T7 promoter ,lac operator and T7 terminator. From its upstream to understream includes StayGold, stem-loop, Twister P1 and mScarlet.

A ribozyme is proved to have cleavage ability when green and red fluorescence are emitted at the same time. We can assess the cleavage efficiency of ribozyme based on the ratio of red-green fluorescence intensity when the stem-loop is unchanged.

We can also assess stem-loop’s ability of preventing mRNA degradation based on the ratio of red-green fluorescence intensity when the ribozyme is unchanged.

Furthermore, the orginal T7_RBS can be replaced by other RBSs. We can test the strength of one RBS based on ratio of red-green fluorescence intensity.

Usage and Biology

This composite part is an easy and effective tool to select the fit ribozyme and stem-loop.

characterization

Sequence and Features