Pakistan Journal of Biological Sciences1028-88801812-5735Asian Network for Scientific Information10.3923/pjbs.1998.179.182Escherichia Coli]]>RajokaM.I. BashirA. 3199813Genes for endo-glucanase (Egl) isolated from a genomic library of the cellulolytic bacterium, Cellulomonas biazotea, were cloned in pUC18 in its Sac1 cloning site and transformed to E. coli. Six clones showed yellow zones of hydrolysis on carboxy methyl cellulose (CMC) plates with Congo red, in liquid culture, and on native polyacrylamide gel electrophoresis activity gels. They belonged to three distinctly different groups. Three representative E. coli clones carrying recombinant palsmids were designated pRM2, pRM13 and pRMC28. The genes were located on 1.1, 2.1 and 4.6 kb fragments respectively. Their location was obtained by deletion analysis which revealed that 0.65, 1.1 and 3.2 kb fragments were essential to code for EglA, EglB and EglC, respectively and conferred intracellular production of endoglucanase on E. coli. Expression of the Egl genes resulted in hyperproduction of endo-glucanase in all clones. The secretion occurred into the periplasmic fractions. The endo-glucanases produced by recombinant E. coli resemble the native endo-glucanases with respect to temperature optima, pH optima, and effect of metal ions on enzyme activities but differ with respect to location of the enzyme and some other enzyme properties. The cloned genes can be used as selection markers for introducing recombinant plasmids in wild strains of E. coli. ]]>Akhtar, M.W., M. Duffy, B.C.A. Dowds, M.C. Sheehan and D.J. McConnell,1988Cellulomonas flavigena encoding endo-β-1,4-glucanases (CM-cellulases).]]>74549553Ali, B.R.S., M.P.M. Romaniec, G.P. Hazlewood and R.B. Freedman,1995Clostridium thermocellum cellulosomes.]]>17705711Ausubel, F.M., R. Brent, R.E. Kingston, D.D. Moore, J.G. Seidman, J.A. Smith and K. Struhl,1996Barron, A., G. May, E. Bremer and M. Villarejo,1986Escherichia coli.]]>167433438Bradford, M.M.,197672248254Brestic-Goachet, N., P. Gunasekaran, B. Cami and J.C. Baratti,1989Erwinia chrysanthemi cellulase gene in Zymomonas mobilis.]]>135893902Esterbauer, H., W. Steiner, I. Labudova, A. Hermann and M. Hayn,1991Trichoderma cellulase in Laboratory and pilot scale.]]>365156Gadgil, N.J., H.F. Oaginawala, T. Chakarbartu and P. Khanna,1995Trichoderma reesei.]]>17942946Gilkes, N.R., D.G. Kilburn, R.C. Miller Jr. and R.A.J. Warren,1991362135Kim, J.H. and M.Y. Pack,1993Bacillus subtilis.]]>15133136Kim, J.M., I.S. Kong and J.H. Yu,1987Bacillus sp. and its expression in Escherichia coli.]]>5326562659Lejeune, A., S. Courtois and C. Colson,1988Pseudomonas fluorescens subsp. cellulosa produced in Escherichia coli and regulation of the expression of its cloned gene.]]>54302308Lindner, W.A.,1988Sclerotium rolfsii.]]>160376382Moser, B., N.R. Gilkes, D.G. Kilburn, R.A. Warren and R.C. Miller,1989Cellulomonas fimi, cloning of the gene and analysis of in vivo transcripts of the gene.]]>5524802487Nakamura, K., N. Misawa and K. Kitamura,1986Cellulomonas uda CB4. I. Cloning and expression of β-glucosidase genes in Escherichia coli.]]>3239246Rajoka, M.I., S. Parvez and K.A. Malik,1992Cellulomonas biazotea into E. coli and Saccharomyces cerevisiae using shuttle vector pBLU-D.]]>1410011006Rajoka, M.I. and K.A. Malik,1997Cellulomonas strains grown on different cellulosic residues.]]>425964Rajoka, M.I., A. Bashir, S.R.S. Hussain and K.A. Malik,1998Cellulomonas biazotea for improved production of cellulases.]]>431522Rajoka, M.I., A. Bashir, S.R.A. Hussain, M.T. Ghauri, R. Parvez and K.A. Malik,1998Escherichia coli and Saccharomyces cerevisiae using shuttle vector pYES 2.0.]]>43129135Sami, A.J. and M.W. Akhtar,1993Cellulomonas flavigena.]]>15586592Siddiqui, K.S., M.J. Azhar, M.H. Rashid and M.I. Rajoka,1997Aspergillus niger and Cellulomonas biazotea.]]>42312318Su, P., C.Q. Liu, R.J. Lucas, S.F. Delaney and N.W. Dunn,1993Zymomonas mobilis.]]>15979984Teather, R.M. and P.J. Wood,198243777780