A cosmid vector that facilitates restriction enzyme mapping.
نویسندگان
چکیده
منابع مشابه
Restriction mapping of retroviral vector episomal DNA.
The infidelity of reverse transcription enzyme activity during the replication cycle of a retrovirus can cause retroviruses to rapidly mutate. For gene therapy, this raises biosafety issues, such as the deletion of a therapeutic gene from a vector and the development of a replication-competent retrovirus. In a single retroviral replication cycle, a therapeutic herpes simplex virus thymidine kin...
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We have designed cosmid vectors for rapid genomic "walking" and restriction mapping. These vectors contain the transcription promoters from either bacteriophage SP6, T7, or T3 flanking a unique BamHI cloning site. Mammalian expression modules encoding the dominant marker neomycin phosphotransferase or the amplifiable dihydrofolate reductase gene expressed from SV40 promoters were inserted for u...
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We describe a simple method for the cloning of PCR products without the need for post-amplification enzymatic treatment. Tailed PCR primer sets are used to create complementary staggered overhangs on both insert and vector by a post-PCR denaturation-hybridisation reaction. The single-stranded overhangs are designed to allow directional cloning in a ligase-free manner. This 'enzyme-free cloning'...
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Many commercially available systems provide a variety of strategies for high-level expression of recombinant proteins. While the prokaryotic and yeast expression systems are highly efficient and easy to use, expression systems for use in higher eukaryotic cells are still hampered by difficulties such as inefficient posttranslational processing, insufficient levels of protein production, restric...
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In order to generate contiguous cosmid coverage of the genome of the protozoan parasite Trypanosoma cruzi for large-scale sequence analysis, a cosmid library of 36864 individual, primary clones was generated. Total genomic DNA of the reference strain CL Brener was fragmented both by partial digestion with MboI and by physical shearing. For cloning, a modified cosmid vector was used that simplif...
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ژورنال
عنوان ژورنال: Proceedings of the National Academy of Sciences
سال: 1985
ISSN: 0027-8424,1091-6490
DOI: 10.1073/pnas.82.10.3159