PA1/04/03-RBS400-SE-B1006-J23118-RBSII-IMM-rrnB T1

Description

This is a combination of SE and IMM into device, that is used to regulate the expression of SE and IMM.

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal EcoRI site found at 2524
12
INCOMPATIBLE WITH RFC[12]
Illegal EcoRI site found at 2524
Illegal NheI site found at 2612
Illegal NheI site found at 2635
21
INCOMPATIBLE WITH RFC[21]
Illegal EcoRI site found at 2524
Illegal BglII site found at 2229
Illegal BamHI site found at 1801
23
INCOMPATIBLE WITH RFC[23]
Illegal EcoRI site found at 2524
25
INCOMPATIBLE WITH RFC[25]
Illegal EcoRI site found at 2524
Illegal NgoMIV site found at 1600
Illegal AgeI site found at 1690
1000
COMPATIBLE WITH RFC[1000]

2021 SZPT-China

Biology

This is a combination of SE (BBa_K3740042) and IMM (BBa_K3740034) into device, that is used to regulate the expression of SE and IMM.

Usage

We connected PA1/04/03-RBS400-SE-B1006（BBa_K3740056）and J23118-RBSII-IMM-rrnB T1 (BBa_K3740053) to the expression vector pSEVA331 by standard assembly, and introduced the connection mixture into G. hansenii ATCC 53582. when the engineering bacteria were lysed, the SE protein can be released to kill Pseudomonas aeruginosa specifically.

Characterization

1. Verification of SE protein antibacterial performance of composite parts

It can be seen from Figure 1(a) that PA1/04/03-RBS300-SE-B1006-J23118-RBSII-IMM-rrnB T1-pSEVA331-G. hansenii ATCC 53582 has no inhibitory effect on PAO1, and the zone of inhibition in (b) is not obvious.

Figure 1,（a）Supernatants from different bacterial strains lysate inhibited the growth of PAO1. (b) Inhibition zone experiment. (c) Strains are used for Figure(a) and Figure(b)