Transformation of the moss Physcomitrella patens using T-DNA mutagenesis.
نویسندگان
چکیده
In this protocol, the transformation of moss (Physcomitrella patens) protoplasts is performed via Agrobacterium-mediated transfer of T-DNA. Protoplasts are incubated with Agrobacterium and acetoseringone in regeneration medium. They are then washed and plated on antibiotic-containing medium to select for T-DNA insertion in stable transformants. The transformation rate for this protocol is typically 10 (expressed as the frequency of stable transformants among regenerants surviving the transformation procedure).
منابع مشابه
Isolation and regeneration of protoplasts of the moss Physcomitrella patens.
This method is adapted from a protocol described by Grimsley et al. (1977). For more information about P. patens as a model organism, see The Moss Physcomitrella patens: A Novel Model System for Plant Development and Genomic Studies (Cove et al. 2009a). For details about the growth of P. patens on cellophane overlay plates, see Culturing the Moss Physcomitrella patens (Cove et al. 2009b). For p...
متن کاملTransformation of the moss Physcomitrella patens using direct DNA uptake by protoplasts.
This protocol describes how to transform moss (Physcomitrella patens) protoplasts using polyethylene glycol (PEG)-mediated DNA uptake. The transformation rates for direct uptake by protoplasts of DNA with and without genomic sequence (a targeting construct) are typically 10 and 10, respectively. (These are the frequencies of stable transformants among regenerants surviving the transformation pr...
متن کاملA new moss genetics: targeted mutagenesis in Physcomitrella patens.
The potential of moss as a model system to study plant biology is associated with their relatively simple developmental pattern that nevertheless resembles the basic organization of the body plan of land plants, the direct access to cell-lineage analysis, their similar responses to plant growth factors and environmental stimuli as those observed in other land plants, and the dominance of the ga...
متن کاملTagged mutagenesis and gene-trap in the moss, Physcomitrella patens by shuttle mutagenesis.
The moss, Physcomitrella patens has been used as a useful material in many fields, because of its simple body plan, ease of gene targeting, and other reasons. Although many mutants have been reported, no method to isolate the corresponding genes was reported. We developed a gene tagging and gene-trap system in P. patens by using the shuttle mutagenesis technique, which has been used in the budd...
متن کاملTransformation of moss Physcomitrella patens gametophytes using a biolistic projectile delivery system.
RELATED INFORMATION For more information about P. patens as a model organism, see The Moss Physcomitrella patens: A Novel Model System for Plant Development and Genomic Studies (Cove et al. 2009a). The growth of protonemal tissue is described in Culturing the Moss Physcomitrella patens (Cove et al. 2009b), and a method for isolation of P. patens protoplasts is found in Isolation and Regeneratio...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Cold Spring Harbor protocols
دوره 2009 2 شماره
صفحات -
تاریخ انتشار 2009