Team:Brown/Parts/In Progress

1 The R Gene - The second gene of the Cell Lysis Cassette. It is an endolysin causing lysis of the bacterial cell. The gene is 477 base pairs in length.

2 The Rz Gene The third gene in the Cell Lysis Cassette. It is 462 base pairs, however, little is known of its function in cell lysis. It is included in many cell lysis methods but has yet to be characterized sufficiently.

Composite Parts

Mercury Sensor BBa_I728456 (Mercury Inducible Promoter) coupled with BBa_K124003 (PVJ4 Cell Lysis Cassette). The current Arabinose Inducible Promoter is replaced with a Mercury Promoter. In the presence of Mercury in a water sample, the three lysis genes, S, R, and Rz genes will be expressed.

Xylene Sensor BBa_I723020 (Pu) and BBa_K124003 (PVJ4). Transcription activated in the presence of environmental xylene.

Arsenic Sensor BBa_J33201 (E. coli chromosomal ars promoter with arsR repressor gene) and BBa_K124003 (PVJ4). A method for detection of Arsenic in water supplies.

LacI BBa_R0010 and BBa_K124003 (PVJ4). When lactose or IPTG is present, lacI is unable to bind and prevent transcription.

T7 BBa_I719005 and BBa_K124003. Part BBa_I719005 is the T7 promoter. Proteins are expressed at high levels in the presence of T7 Polymerase.

The AND Gate

The individual S and R genes can be placed under different promoters, creating an AND Gate. For example, the S gene can be placed under an Arabinose promoter while the R gene is placed under an IPTG (isopropyl-beta-D-thiogalactopyranoside) promoter. (The IPTG promoter induces gene expression of genes under the control of the lac operon.) The S and R genes will not be expressed unless both "toxins" are present. If one of the toxins is present and the other is not, there will be little to no gene expression of the Lysis genes.