Translocation of double stranded DNA through membrane adapted phi29 motor protein nanopore
نویسندگان
چکیده
Biological pores have been used to study the transport of DNA and other molecules, but most pores have channels that allow only the movement of small molecules and single-stranded DNA and RNA. The bacteriophage phi29 DNA-packaging motor, which allows double-stranded DNA to enter the virus during maturation and exit during an infection, contains a connector protein with a channel that is between 3.6 and 6 nm wide. Here we show that a modified version of this connector protein, when reconstituted into liposomes and inserted into planar lipid bilayers, allows the translocation of double-stranded DNA. The measured conductance of a single connector channel was 4.8 nS in 1 M KCl. This engineered and membrane-adapted phage connector is expected to have applications in microelectromechanical sensing, microreactors, gene delivery, drug loading and DNA sequencing.
منابع مشابه
Microscopic Kinetics of DNA Translocation through synthetic nanopores.
We have previously demonstrated that a nanometer-diameter pore in a nanometer-thick metal-oxide-semiconductor-compatible membrane can be used as a molecular sensor for detecting DNA. The prospects for using this type of device for sequencing DNA are avidly being pursued. The key attribute of the sensor is the electric field-induced (voltage-driven) translocation of the DNA molecule in an electr...
متن کاملInvestigating asymmetric salt profiles for nanopore DNA sequencing with biological porin MspA
Nanopore DNA sequencing is a promising single-molecule analysis technology. This technique relies on a DNA motor enzyme to control movement of DNA precisely through a nanopore. Specific experimental buffer conditions are required based on the preferred operating conditions of the DNA motor enzyme. While many DNA motor enzymes typically operate in salt concentrations under 100 mM, salt concentra...
متن کاملDetecting translocation of individual single stranded DNA homopolymers through a fabricated nanopore chip.
Fabricated solid-state nanopore chips are used to probe individual single stranded DNA (ssDNA) homopolymers. Based on the analysis of the current blockage caused by DNA translocation through a voltage-biased nanopore, we discovered that the hydrodynamic diameter of ssDNA homopolymer helix is comparable to that of double stranded DNA (dsDNA) helix. This proof-of-principle demonstration shows tha...
متن کاملRobust properties of membrane-embedded connector channel of bacterial virus phi29 DNA packaging motor.
Biological systems contain highly-ordered macromolecular structures with diverse functions, inspiring their utilization in nanotechnology. A motor allows linear dsDNA viruses to package their genome into a preformed procapsid. The central component of the motor is the portal connector that acts as a pathway for the translocation of dsDNA. The elegant design of the connector and its channel moti...
متن کاملOne-Way Traffic of a Viral Motor Channel for Double-Stranded DNA Translocation
Linear double-stranded DNA (dsDNA) viruses package their genome into a procapsid using an ATP-driven nanomotor. Here we report that bacteriophage phi29 DNA packaging motor exercises a one-way traffic property for dsDNA translocation from N-terminal entrance to C-terminal exit with a valve mechanism in DNA packaging, as demonstrated by voltage ramping, electrode polarity switching, and sedimenta...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
دوره 4 شماره
صفحات -
تاریخ انتشار 2009