EMAIL THIS PAGE TO A FRIEND

Methods in molecular biology (Clifton, N.J.)

Improved vectors for selection of transgenic Caenorhabditis elegans.


PMID 23104336

Abstract

The generation of transgenic animals is an essential part of research in Caenorhabditis elegans. One technique for the generation of these animals is biolistic bombardment involving the use of DNA-coated microparticles. To facilitate the identification of transgenic animals within a background of non-transformed animals, the unc-119 gene is often used as a visible marker as the unc-119 mutants are small and move poorly and the larger size and smoother movement of rescued animals make them clearly visible. While transgenic animals can be identified from co-bombardment with a transgene of interest and a separate unc-119 rescue plasmid, placing the unc-119 in cis on the transgene increases confidence that the resulting transgenic animals contain and express both the marker and the transgene. However, placing the unc-119 marker on the backbone of a plasmid or larger DNA construct, such as a fosmid or BAC, can be technically difficult using standard molecular biology techniques. Here we describe methods to circumvent these limitations and use either homologous recombination or Cre-LoxP mediated recombination in Escherichia coli to insert the unc-119 marker on to a variety of vector backbones.

Related Materials

Product #

Image

Description

Molecular Formula

Add to Cart

R6142 Acc I from Acinetobacter calcoaceticus, Restriction Enzyme
R6885 Alu I from Arthrobacter luteus, Restriction Enzyme
R8631
Bcl I from Bacillus caldolyticus, Restriction Enzyme
R6377
Bgl II from Bacillus licheniformis, Restriction Enzyme
R3131
Bln I from Brevibacterium linens, buffered aqueous glycerol solution
R3635
Bsm I from Bacillus stearothermophilus NUB 36, Restriction Enzyme
R4253 BstE II from Bacillus stearothermophilus ET, Restriction Enzyme
R1761
Cfo I from Clostridium formicoaceticum, Restriction Enzyme
R7763
Cla I from Caryophanon latum L, Restriction Enzyme
R4256
Dde I from Desulfovibrio desulfuricans strain Norway, Restriction Enzyme
R4381
Dra I from Deinococcus radiophilus, Restriction Enzyme
R3884 EclX I from Enterobacter cloacae 590, Restriction Enzyme
R6265
EcoR I from Escherichia coli BS5, Restriction Enzyme
R2756
EcoR V from Escherichia coli, buffered aqueous glycerol solution
R5628 Hae III from Haemophilus aegyptius, Restriction Enzyme
R1632
Mva I from Micrococcus varians Rfl 19, Restriction Enzyme
R8761 Nco I from Nocardia corallina, Restriction Enzyme
R5634
Nhe I from Neisseria mucosa heidelbergensis, Restriction Enzyme
R8506
Not I from Nocardia otidiscaviarum, Restriction Enzyme
R5884 Nsi I from Neisseria sicca, Restriction Enzyme
R7023
Pst I from Providencia stuartii, Restriction Enzyme
R1508
Pvu I from Proteus vulgaris, Restriction Enzyme
R2631
Pvu II from Proteus vulgaris, Restriction Enzyme
R4756 Rsa I from Rhodopseudomonas sphaeroides, Restriction Enzyme
R0754 Sal I from Streptomyces albus G, Restriction Enzyme
R8256
Sfi I from Streptomyces fimbriatus, Restriction Enzyme
R7135 Sph I from Streptomyces phaeochromogenes, Restriction Enzyme
R7260
Xba I from Xanthomonas badrii, Restriction Enzyme
R6379
Xho I from Xanthomonas holcicola, Restriction Enzyme
R1259 Afl III from Anabaena flos-aquae, buffered aqueous glycerol solution