Difference between revisions of "Part:BBa K2230002"

Revision as of 04:35, 25 October 2017

RBS-EmR/pSB1A2

Erythromycin Resistance Gene as a Selection Marker for Lactobacillus transformation

Erythromycin is a common selection marker for engineering lactic acid bacteria1. And teams Uppsala and TMMU_China were using it to select the engineered Lactobacillus with erythromycin resistant strains. However, the BioBrick parts containing the erythromycin resistance gene are currently unavailable. On the other hand, iGEM HQ is unable to service the request of plasmid carrying erythromycin resistance gene. Maybe iGEM HQ can’t manipulate erythromycin resistant bacteria so far. Therefore, we decided to clone the erythromycin resistance gene (EmR) by ourselves. We searched the vector carrying EmR and found an iGEM team HKUST in 2010 working with it. We got the pMG36e vector from the team and cloned the EmR gene out onto pSB1C3. We also made RBS-EmR/pSB1C3 (Bba_K2230002) and RBS-EmR-TT/pSB1C3 (Bba_K2230002) by assembling RBS (B0034) and double terminator (B0015) parts, respectively. These will provide iGEM team this resistance gene as selection marker in the future.

Cloning

EmR (Erythromycin resistance gene) was amplified from pMG36e vector and cut by XbaI & PstI. Then, the fragment was assembled with RBS/pSB1A3 (BBa_B0034) cut by SpeI & PstI

Reference

Genetic engineering techniques for lactic acid bacteria: construction of a stable shuttle vector and expression vector for β-glucuronidase. Biotechnol Lett. 2014;36(2):327-35.

Sequence and Features