生物实验室材料 方法大全pE194ts

2. Materials and Methods

2.1. Enzymes and Chemicals

Amersham-Pharmacia, Braunschweig ECL random primer and labelling kit, Hybond-N+,

Hyperfilm ECL, SureClone ligation kit, Restriction

enzymes, 35S-Methionine in vivo labelling grade,

Megaprimer DNA labelling systems, (α-32P)dCTP. Roche, Mannheim Agarose, PCR-nucleotide mix, Expand long

template PCR-kit, restriction enzymes, Taq-DNA

polymerase, T4-DNA ligase, T4-DNA polymerase,

protease-inhibitor set.

Carl Roth, Karlsruhe Ampicillin, BSA, Chloramphenicol, EDTA, Acetic

acid, Ethanol, Ethidiumbromid, Fructose, Glucose,

Glycerin, HCl, IPTG, MgCl2, Sodium acetate, NaCl,

nButanol, Phenol, Phenol-Chloroform, Proteinase K,

Sucrose, SDS, Tris-HCl, X-Gal, rotiphorese Gel30

(30% acrylamid, 0.8% bisacrylamid solutions).

Difco Laboratories, Augsburg Agar-Agar, Trypton broth, Yeast extract.

Merck, Darmstadt KH2PO4, K2HPO4.

Millipore, Eschborn Nitrocellulose (0,025 µm) .

New England-Biolabs, Schwalbach/Ts. Restriction enzymes, Shrimp alkali phosphatase. Qiagen, Hilden Plasmid Midi kit (50), Plasmid Maxi kit (10),

QIAEx gel extraction kit, QIAquick PCR

purification kit, QIAGEN Genomic-tip 100/G and

500/G, Rneasy Mini kit, Ni-NTA superflow (25),

His Antibody (100).

Promega, pGEM-T vector systems.

Sartorius, Göttingen Sterilfilter (0.2 µm).

Sigma-Aldrich, Steinheim ATP, MnSO4, Papain, Pepsin, PMSF, amino acids. Serva,

Heidelberg Succinat, Dextransulfat, Dimethylsulfoxid,

Lysozyme, RNase, Triton X-100, Tween20. Winthorp, Dublin, Ireland Kanamycin.

2.2. Strains and growth conditions

2.2.1. Strains

Table 1: Bacterial strains

Bacteria Geno-/Phenotype & Reference

Bacillus amyloliquefaciens

GBA12 ALKO2718; ∆nprE, ∆aprE;Vehmaanperrä, et al.,

(1991)

but

sipS::pEAS* a; This study

GBA13 GBA12,

sipT::pEAT*; This study

GBA14 GBA12,

but

but sipV::pEAV*; This study

GBA15 GBA12,

sipW::pEAW*; This study

but

GBA16 GBA12,

15841

Bacillus amyloliquefaciens ATCC

Bacillus amyloliquefaciens ATCC 23350 = DSM 7T

Bacillus amyloliquefaciens ATCC

23842

Bacillus amyloliquefaciens IAM 1523 DSM No. 1061

Bacillus amyloliquefaciens IFO 3034 DSM No. 1062

Bacillus amyloliquefaciens IFO 3037 DSM No. 1063

Bacillus amyloliquefaciens KA 63 DSM No. 1060

Bacillus amyloliquefaciens OUT 8419 DSM No. 1064

Bacillus amyloliquefaciens OUT 8420 DSM No. 1065

Bacillus amyloliquefaciens OUT 8421 DSM No. 1066

Bacillus amyloliquefaciens OUT 8426 DSM No. 1067

Bacillus amyloliquefaciens N strain collection of Ag BAG, IPK

Bacillus amyloliquefaciens P strain collection of Ag BAG, IPK

Bacillus amyloliquefaciens SB I strain collection of Ag BAG, IPK

Bacillus amyloliquefaciens T strain collection of Ag BAG, IPK

Bacillus amyloliquefaciens ZFL 14/4strain collection of Ag BAG, IPK

Bacillus amyloliquefaciens ZF 178 strain collection of Ag BAG, IPK

Bacillus brevis 475 Q strain collection of Ag BAG, IPK

Bacillus circulans GB2 strain collection of Ag BAG, IPK

Bacillus lentus 3601 FZB strain collection of Ag BAG, IPK

Bacillus licheniformis 41p strain collection of Ag BAG, IPK

Bacillus macerans B30 strain collection of Ag BAG, IPK