Cloned gene expression

If the end goal of cloning is to have a cloned gene expressed in a cell, the entire coding sequence must be doned intact. Furthermore, if a cloned eukaryotic gene is to be expressed in bacteria (to make recombinant proteins), the gene must not contain introns, which could not be processed in a prokaryotic cell. In these cases it is more convenient to done cDNA rather than DNA restriction fragments. 

Fujiwara, H. et al., cDNA cloning, gene expression and subcellular localization of anthocyanin 5-aromatic acyltransferase from Gentiana triflora. Plant J., 16, 421, 1998. 

Xgtll Bacteriophage E. coli cDNA cloning, gene expression... 

XZAP Bacteriophage E. coli Resistance to ampicillin. lac screening. cDNA cloning, gene expression, automatic excision into plasmid pBluescript SK... 

Mattsson JP, Li X, Peng S-B, Nilsson F, Adersen P, Lundberg LG, Stone DK, Keeling DJ. 2000. Properties of three isoforms of the 116-kDa Subunit of vacuolar H+-ATPase from a single vertebrate species. Cloning, gene expression and protein characterization of functionally distinct isoforms in Gallus domesticus. Euro J Bioch 267 4115-26. 

FUJIWARA, H., TANAKA, Y., YONEKURA-SAKAKIBARA, K., FUKUSHI-MIZUTANI, M., NAKAO, M FUKUI, Y., YAMAGUCHI, M., ASHIKARA, T., KUSUMI, T., cDNA cloning, gene expression, and subcellular localization of anthocyanin 5-aromatic acyltransferase from Gentiana triflora. Plant J., 1998,16, 421-431. 

Robertus J, Piatak M, Ferris R, Houston L. Crystallization of ricin A chain obtained from a cloned gene expressed in Escherichia coli. J Biol Chem. 1987 262 19-20. 

Methods for rapid gene cloning Gene expression Mialysis (DNA arrays, Northern s)... 

Key words Gene synthesis. Codon optimization. Polymerase chain reaction. Cloning, Gene expression. Oligonucleotide design. Simulated annealing... 

Subunit vaccines based on the surface proteins of vims are also being explored. It has been demonstrated that the two major protective antigens are haemagglutinin (HA) and neuraminidase (NA). The genes for these antigens have been cloned and expressed in baculovims in insect cell culture (84). 

Miyamoto, C., Boylan, M., Graham, A., and Meighen, E. (1986). Cloning and expression of the genes from the bioluminescent system of marine bacteria. Method. Enzymol. 133 70-83. 

Cell-free translation system, used for the identification of cloned genes and gene expression, has been investigated extensively as a preparative production system of commercially interesting proteins after the development of continuous-flow cell-free translation system. Many efforts have been devoted to improve the productivity of cell-free system [1], but the relatively low productivity of cell-free translation system still limits its potential as an alternative to the protein production using recombinant cells. One approach to enhance the translational efficiency is to use a condensed cell-free translation extract. However, simple addition of a condensed extract to a continuous-flow cell-free system equipped with an ultrafiltration membrane can cause fouling. Therefore, it needs to be developed a selective condensation of cell-free extract for the improvement of translational efficiency without fouling problem. 

Once a gene is cloned it is necessary to convert the information contained in it into a functional protein. There are a number of steps in gene expression (i) transcription of DNA into mRNA (ii) translation of the mRNA into a protein sequence and (iii) in some instances, post-translational modification of the protein. In discussing these steps in more detail, expression of a cloned insulin gene will be used as an example. 

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