By Rocky S. Tuan
ISBN-10: 0896033333
ISBN-13: 9780896033337
Professional overseas examine employees supply the main updated and complete choice of thoughts on hand for expressing a number of genes of curiosity. The authoritative individuals describe extensive the vectors used for expression in micro organism, yeast, viruses, nonmammalian larger eukaryotes, mammals, and crops, in addition to in transgenic animals. in addition they supply worthwhile specified info on find out how to introduce the recombinant gene build into definitely the right expression host. whilst mixed with a better half quantity via an analogous editor, Recombinant Proteins: Detection and Isolation Protocols, either volumes consultant the study employee during the whole trip of recombinant gene expression.
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Sample text
2. Promoter induction The two promoters most often used m h expression systems are the E. colz lad and bacteriophage T7 gene 10 promoters. The 1acZ promoter may be leaky, particularly if the host cell has a single copy lad gene and the 1acZ promoter is multicopy. The Ia& mutation overexpresses the lac repressor, thus maintaining more efficient repression of the 1acZ promoter. The 1acI gene can also be put on a multicopy plasmid to decrease the background level of expression from 1ucZ The 1acZ gene is typically derepressed by addition of IPTG Since IPTG is not hydrolyzed by P-galactosidase, derepression is continuous once the IPTG has been added.
Capon and coworkers (45) demonstrated that by fusing a soluble portton of CD4 (the target receptor for the human nnmunodeficiency vnus particle) to the Fc part of IgG, the serum half-life of CD4 m rabbits increased 200-fold. Nygren and coworkers (46) described an alternattve strategy where in vtvo stabthzation was achieved by fusion of the unstable protein to a bacterial receptor capable of bmdmg to serum albumms. The serum half-lives of different fusion proteins containing the albumin-bmdmg receptor 44 SkihI, Nygren, and Uhlbn were tested m mice.
15 Obukowtcz, M G , Turner, M A , Wong, E Y , and Tacon, W C (1988) Secretion and export of IGF- 1 m Escherzchla colz strain JMI 01. A401 Gen Genet 215, 19-25. 16. , Gussow, D , Grtffiths, A D , Jones, P. T , and Winter, G. , Perlmann, P , and Wahlgren, M (1993) Plasmodlumfalclparum The immune response m rabbits to the clustered asparagme-rich protein (CARP) after nnmuntzatton m Freund’s admvant or rmmunosttmulatmg complexes (ISCOMs) Exp Parasztol 76, 134-145 24 Rudolph, R (1995) Successful protein foldmg on an mdustrial scale, m Prznczples andpractzce ofProtezn Foldzng (Cleland,J L and Cratk, C.
Recombinant Gene Expression Protocols by Rocky S. Tuan
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