NEW CHROMOSOMAL JUMPING VECTORS FOR GENE MAPPING

CHROMOSOMAL JUMPING HAS BEEN PROPOSED AS A RAPID TECHNIQUE FOR ORDERING LARGE SEGMENTS OF THE GENOME, INCREASING THE SPEED AT WHICH GENES ASSOCIATED WITH HEREDITABLE DISEASES INHUMANS CAN BE ISOLATED. TECHNICAL DIFFICULTIES ASSOCIATED WITH THE CONSTRUCTION OF THE NECESSARY JUMPING AND JUNCTION LIBRARIES HAVE SLOWED THE SUCCESSFUL APPLICATION OF THIS TECHNIQUE. THE MOST DIFFICULT STEP IN THE CONSTRUCTION OF JUMPING LIBRARIES IS CIRCULARIZING EXTREMELY LARGE GENOMIC DNA FRAGMENTS ONTO A SELECTABLE MARKER IN A SINGLE REACTION. STRATAGENE CLONING SYSTEMS WILL DEVELOP A NOVEL CLONING TECHNOLOGY THAT ALLOWS THIS STEP TO BE REDUCED TO A SIMPLE CIRCULARIZATION WITHOUT THE SELECTABLE MARKER. THE SELECTABLE MARKER IS ADDED AT A LATER STEP UNDER CONDITIONS THAT HIGHLY FAVOR THE DESIRED LIGATION PRODUCTS. THIS TECHNOLOGY HAS THE ADDITIONAL FEATURE OF T3 AND T7 RNA POLYMERASE PROMOTERS POSITIONED TO GENERATE HIGH-SPECIFIC-ACTIVITY PROBES WITHOUT SUBCLONING OR FRAGMENTISOLATION.