PA1/04/03-RBS150-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 2528
12
INCOMPATIBLE WITH RFC[12]
Illegal EcoRI site found at 2528
Illegal NheI site found at 2616
Illegal NheI site found at 2639
21
INCOMPATIBLE WITH RFC[21]
Illegal EcoRI site found at 2528
Illegal BglII site found at 2233
Illegal BamHI site found at 1805
23
INCOMPATIBLE WITH RFC[23]
Illegal EcoRI site found at 2528
25
INCOMPATIBLE WITH RFC[25]
Illegal EcoRI site found at 2528
Illegal NgoMIV site found at 1604
Illegal AgeI site found at 1694
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-RBS150-SE-B1006（BBa_K3740054）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, SE protein can be released to kill Pseudomonas aeruginosa specifically.

Figure 1. Gene circuit of PA1/04/03-RBS150-SE-B1006-J23118-RBSII-IMM-rrnB T1.

Characterization

1. Verification of SE protein antibacterial performance of composite parts

As shown in (a), the lysis supernatant of PA1/04/03-RBS150-SE-B1006-J23118-RBSII-IMM-rrnB T1-pSEVA331-G. hansenii ATCC 53582-2# has an inhibitory effect on the growth of PAO1; In (b), the lysis supernatant produced a zone of inhibition, but the effect was not very obvious. These results show that expression of SE protein was successfully induced in G. hansenii ATCC 53582 and indicated that PA1/04/03-RBS150-SE-B1006-J23118-RBSII-IMM-rrnB T1-pSEVA331-G. hansenii ATCC 53582-2# has an inhibitory effect on PAO1, but the effect is not very obvious.

Figure 2.（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)