Gene Transfer and Expression: A Laboratory Manual by Michael Kriegler (auth.)

By Michael Kriegler (auth.)

A sensible handbook of protocols for attaining expression of overseas genes in mammalian cells. It contains a few very new innovations resembling PCR-based expression. the writer supplies a theoretical advent to the protocols and compares the strengths and weaknesses.

Show description

Read or Download Gene Transfer and Expression: A Laboratory Manual PDF

Best methodology & statistics books

Image and Reality: Kekule, Kopp, and the Scientific Imagination

Nineteenth-century chemists have been confronted with a specific challenge: tips on how to depict the atoms and molecules which are past the direct succeed in of our physically senses. In visualizing this microworld, those scientists have been the 1st to maneuver past high-level philosophical speculations in regards to the unseen. In snapshot and truth, Alan Rocke specializes in the group of natural chemists in Germany to supply the root for a fuller figuring out of the character of medical creativity.

Readings in Social Research Methods, 3rd Edition

This reader is designed to accompany any major textual content in examine tools or be used as a stand-alone reader. it's been heavily patterned at the diversity of themes coated in Earl Babbie's most sensible promoting texts, THE perform OF SOCIAL examine, 11/e and fundamentals OF SOCIAL examine, 4/e. The reader specializes in the center methodologies of the social examine equipment path and offers illustrations of these equipment.

Introductory Mass Spectrometry, Second Edition

Mass spectrometry has performed a vital part within the research of natural molecular constructions for greater than 50 years, supplying major details from small quantities of pattern. The mass spectrum produced by means of electron impression ionization offers a development of peaks which could usually provide definitive structural information regarding an unknown compound.

Additional info for Gene Transfer and Expression: A Laboratory Manual

Example text

ADENOVIRUS-BASED VECTORS Unfortunately, the development of such cell lines has not led to the development of expression vectors as sophisticated as those described for SV40-based COS transfection. The result of such neglect is that COS-based transfection is the overwhelming favorite for transiently expressing genes at high levels for purposes such as expression cloning. Nevertheless, the MOP and WOP cell lines may be appropriate for your use, particularly if you intend to construct your own polyoma virus expression vector.

To address this problem, a generalized adenovirus vector system was developed. The vector carried a complete structural gene encoding the SV40 T antigen whose expression was driven by its own promoter. The foreign genes of interest, either the polyoma T-antigen genes or the HSVtk gene, were inserted just upstream of the SV40 T-antigen cistron, and just downstream from the adenovirus MLP and tripartite leader. Neither the polyoma T antigens nor the HSVtk gene provides adenovirus helper function.

The gpt-encoding, dominant selectable, vaccinia virus open reading frame vectors described in the text are depicted here. These vectors contain vaccinia virus tk gene sequences flanking the E. 5), and the pUC-derived ampicillin resistance gene and origin of replication. The unique cloning sites into which foreign DNA can be inserted in both plasmids downstream of the initiation codon of the gene encoding the 11K peptide are indicated. Two derivatives of pTKgpt-F1s that shift the reading frame of the inserted sequence one nucleotide (pTKgpt-F2s) and two nucleotides (pTKgpt-F3s) relative to the 11K initiation codon have been generated that permit the insertion of foreign sequences into all three possible reading frames.

Download PDF sample

Rated 4.32 of 5 – based on 32 votes