Choose a vector
c. Request PTF Vectors
- We can provide binary vector pTF101.1 for subcloning of your gene of
interest for plant transformation. Note that the vector is not an expression
vector, therefore, there is no promoter for your gene of interest. pTF101.1
is a suitable for you to subclone your entire gene cassette (promoter+gene+terminator).
- The preferred Agrobacterium strain for pTF101.1 is EHA101 (a
strain harbors a derivative of the agropine/L,L-succinamopine-type
Ti-plasmid pTiBo542).
- Basic features of the vector:
- A broad host range (pVS1) origin of replication.
- An aadA gene for bacterial selection of spectinomycin
resistance.
- The T-DNA right and left border fragments from nopaline strain
of A. tumefaciens.
- A selectable marker gene cassette containing (1) 2x CaMV 35S
promoter, (2) TEV (Tobacco Etch Virus translational enhancer), (3)
bar coding region, (4) nos terminator.
- A multiple cloning site containing seven unique restriction
enzyme sites (Hind III, Xba I, BamH I, Xma I, Sma I, Sac I, EcoR I).
- Types of pTF101.1
- MCS version (map,
components,
sequences,
publication,
MTA)
pTF101.1 contains a multiple cloning site carrying seven unique
restriction enzyme sites (Hind III, Xba I, BamH I, Xma I, Sma I, Sac
I, EcoR I).
- Gateway version
- pTF101.1gw1 (map,
components,
MTAgw) This Gateway one-entry version contains attR1 and
attR2 recombination system suitable for subcloning your
Gateway compatible gene of interest cassette (promoter+gene+terminator).
- pTF101.1gw3 (map,
components,
MTAgw) This Gateway three-entry version contains attR3
and attR4 recombination system suitable for subcloning your
Gateway compatible promoter, gene and terminator fragments
all at once.
- Please follow the steps below for requesting these vectors:
- Down load the standard Material Transfer Agreement (MTA,
MTAgw).
- Complete MTA by filling the information and obtaining signatures
of both Principle Investigator (PI) and the institutional authorized
officer.
- Send your request by e-mail to Kan Wang (kanwang@iastate.edu)
including full mailing address and specific request on desired
vector.
- Send back the signed MTA by fax 515-294-2299 (attention Kan
Wang) or electronic attachment to
kanwang@iastate.edu.
- Send an acknowledgement mail to
kanwang@iastate.edu upon receiving the vector DNA by mail.