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008 161107s2016 ua dh f m 000 0 eng d
040 _aEG-GiCUC
_beng
_cEG-GiCUC
041 0 _aeng
049 _aDeposite
097 _aM.Sc
099 _aCai01.07.10.M.Sc.2016.Na.G
100 0 _aNagwa Mohamed Abdelaziz Aly
245 1 0 _aGenetic studies on keratinase producing bacteria /
_cNagwa Mohamed Abdelaziz Aly ; Supervised Naglaa Abdelmoneim Abdallah , Abdelhadi Abdallah Abdelhadi , Mohamed Safwat Abdelsalam
246 1 5 _akeratinase دراسات وراثية على البكتيريا المنتجة للـ
260 _aCairo :
_bNagwa Mohamed Abdelaziz Aly ,
_c2016
300 _a145 P. :
_bcharts , facsimiles ;
_c25cm
502 _aThesis (M.Sc.) - Cairo University - Faculty of Agriculture - Department of Genetics
520 _aThe main objective of this work is to select and genetically improve keratinase producing bacteria in order to reduce environmental feather pollution and convert it to useful products. Fourty two bacteria were isolated, all were screened for keratinolytic activity. Among those, one showed high keratinase specific activity of 28.88 U/ml, and was identified as Bacillus spp. Morphological, biochemical characteristic, growth and molecular level 16s rRNA indicated 99% similarity with Bacillus subtilis sub spiziziniie. Therefore, it was named Bacillus subtilis sub spiziziniie N23. this isolate was mutagenized using N-methyl-N{u2032}-nitro-N- nitrosoguanidine (NTG) mutagenesis and resulted in a high mutant no.12 as it produced 34.92 U/ml. In addition, five collected Bacillus strains were mutagenized using NTG mutagenesis: Bacillus licheniformis N5 had 40.76 U/ml, resulted in a high mutant no.58 (49.16 U/ml), Bacillus pumilis I1 had 27.76 U/ml, resulted in a high mutant no.17 (39.56 U/ml), Bacillus megaterium 7A37 had 23.80 U/ml, resulted in a high mutant no.7 (52.04 U/ml), Bacillus cereus 6A15 had 20.88 U/ml, resulted in a high mutant no.16 (37.12 U/ml). and Bacillus licheniformis 5A3 had 36.64 U/ml, resulted in a high mutant no.42 that had 44.20 U/ml. The highest mutants in keratinase specific activities were nos.58 and 7 and were used for protoplast fusion trail. Two fusants were grown on selective media. Fusant no.1 had 41.84 U/ml, fusant no.2 had 41.44 U/ml in keratinase specific activities. Keratinase gene from the chromosomal DNA of isolate Bacillus subtilis N23 Mutant no.12 was successfully amplified by PCR using keratinase specific primer, cloned into pGEM- Teasy cloning vector and transferred to Escherichia coli DH5Ü. Sequence analysis showed four transformants, two transformants designated as E.coli DH5Ü pGEM ker Bacillus subtilisN23-1,2 and two transformants designated as E.coli DH5Ü pGEM ker Bacillus subtilisN23-M12-1,2. The ker gene is succesfuly expressed in E. coli DH5Ü. Escherichia coli DH5Ü pGEM ker Bacillus subtilisN23 and Escherichia coli DH5Ü pGEM ker Bacillus subtilisN23-M12 produced extracellular keratinase specific activities of 52.16 and 53.78 U /ml, respectively
530 _aIssued also as CD
653 4 _aFeather
653 4 _aKeratin
653 4 _aKeratinase
700 0 _aAbdelhadi Abdallah Abdelhadi ,
_eSupervisor
700 0 _aMohamed Safwat Abdelsalam ,
_eSupervisor
700 0 _aNaglaa Abdelmoneim Abdallah ,
_eSupervisor
856 _uhttp://172.23.153.220/th.pdf
905 _aEnas
_eCataloger
905 _aNazla
_eRevisor
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_cTH
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