Directional cloning

Chapter 4 review the fundamentals of gene structure and function, we are ready to start a more detailed study of molecular biology. The main focus of chapter will be the experiments that molecular biologists have performed to elucidate the structure and function of genes.

67p tangtuy05 30-03-2016 93 6   Download

We found a leucine aminopeptidase (LAP; EC 3.4.11.1) to be abundant in meiotic prophase tissue of a basidiomycete, Coprinus cinereus. After direct puri®cation of the amino-peptidase component from meiocytes, we cloned the gene by degenerate PCR using partial amino-acid sequences of the puri®ed enzyme and 5¢ and 3¢ RACE. It was homologous to the eukaryotic leucine aminopeptidase gene.

7p research12 01-06-2013 46 6   Download

The relevance of functional amino acids for taurocholate transport by the sodium-dependent taurocholate cotrans-porting polypeptide Ntcp was determined by site-directed mutagenesis. cRNAfrom28 single-pointsmutants of the rat liver Ntcp clone was expressed in Xenopus laevisoocytes.

11p tumor12 20-04-2013 46 5   Download

The extremely heat-stable 5¢-methylthioadenosine phos-phorylase from the hyperthermophilic archaeonPyrococcus furiosuswas cloned, expressed to high levels inEscherichia coli, and purified to homogeneity by heat precipitation and affinity chromatography. The recombinant enzyme was subjected to a kinetic analysis including initial velocity and product inhibition studies. The reaction follows an ordered Bi–Bi mechanism and phosphate binding precedes nucleo-side binding in the phosphorolytic direction....

11p awards 05-04-2013 39 2   Download

In the nonmevalonate pathway of isoprenoid biosynthesis, the conversion of 2C-methyl-D-erythritol 4-phosphate into its cyclic diphosphate proceeds via nucleotidyl intermediates and is catalyzed by the products of theispD,ispE andispF genes. An open reading frame of Campylobacter jejuni with similarity to theispDandispFgenes ofEscherichia coli was cloned into an expression vector directing the formation of a 42 kDa protein in a recombinantE. colistrain.

8p awards 05-04-2013 39 5   Download

Volkensin, a type 2 ribosome-inactivating protein from the roots of Adenia volkensiiHarms (kilyambiti plant) was characterized both at the protein and nucleotide level by direct amino acid sequencing and cloning of the gene enco-ding the protein. Gene sequence analysis revealed that vol-kensin is encoded by a 1569-bp ORF (523 amino acid residues) without introns, withan internal linker sequence of 45 bp.

10p dell39 03-04-2013 67 3   Download

The penicillin-binding proteins (PBPs) are ubiquitous bac-terial enzymes involved in cell wall biosynthesis,and are the targets of theb-lactam antibiotics. The low molecular mass Neisseria gonorrhoeaePBP 4 (NG PBP 4) is the fourthPBP revealed in the gonococcal genome. NG PBP 4 was cloned, overexpressed,purified,and characterized for b-lactam binding,DD-carboxypeptidase activity,acyl-donor substrate specificity,transpeptidase activity, inhibitionby a number of active site directed reagents,and pHprofile....

10p dell39 03-04-2013 37 3   Download

We report the development of a high-level bacterial expression system for the Alzheimer’s disease-associated amyloid b-peptide (Ab), together with a scaleable and inexpensive purification procedure. Ab(1–40) and Ab(1–42) coding sequences together with added ATG codons were cloned directly into a Pet vector to facilitate production of Met-Ab(1–40) and Met-Ab(1– 42), referred to as Ab(L1–40) and Ab(L1–42), respectively.

11p vinaphone15 27-02-2013 48 2   Download

Directed Random Testing bool var35 = Collections.replaceAll(var20, var23, var28); java.lang.Object var36 = var18.put(1L, 10L); int var37 = Arrays.binarySearch(var4, var23); assert var37 == 3; Arrays.fill(var1, var3, var37, -1); BitSet var38 = new BitSet(var37); Comparator var39 = Collections.reverseOrder(); PriorityQueue var40 = new PriorityQueue(var37, var39); TreeSet var41 = new TreeSet(var39); bool var42 = var41.isEmpty(); TreeSet var43 = (TreeSet)var41.clone(); assert var43.size() == var41.size(); char[] var44 = new char[] { }; Set var76 = Collections.

72p quangdat88 17-01-2013 42 3   Download

Tuyển tập các báo cáo nghiên cứu về lâm nghiệp được đăng trên tạp chí lâm nghiệp Original article đề tài: Direct effects of acidic wet deposition on photosynthesis and stomatal conductance of two Populus clones (P. cv. Beaupré and P. cv. Robusta)...

4p toshiba9 12-10-2011 38 1   Download

Norton Ghost protects your data from computer disasters. The intuitive Windows interface lets you create backup images of your hard disk or selected partitions. You can clone directly between two computers using a network, USB, or parallel connection.

221p nguyen5 24-11-2009 101 8   Download