KOSEN 한인과학기술자네트워크

둘다 염기서열이 확실치 않은 것 같은데요... pHPS9는 ATCC (ATCC® Number: 37817) 에서 제공되고 있는 vector인 것 같은데 염기서열 정보가 확실치 않네요... 아마 아래 논문을 참고로 해서 서열을 조합해 봐야할 것 같은데요... 1) Haima P , et al. Development of a beta-galactosidase alpha-complementation system for molecular cloning in Bacillus subtilis. Gene 86: 63-69, 1990. PubMed: 2107125 2) Haima P , et al. An improved beta-galactosidase alpha-complementation system for molecular cloning in Bacillus subtilis. Gene 93: 41-47, 1990. PubMed: 2121609 3) Haima P , et al. Novel plasmid marker rescue transformation system for molecular cloning in Bacillus subtilis enabling direct selection of recombinants. Mol. Gen. Genet. 223: 185-191, 1990. PubMed: 2123518 pHPS9에 관련된 간단한 설명입니다 (http://seq.yeastgenome.org/vectordb/vector_descrip/PHPS9.html) Cloning into the NdeI, NheI, BamHI, SmaI or EcoRI sites inactivates lacZalpha. The NcoI site interrupts the chloramphenicol resistance sequence. [1] Permits alpha-complementation to identify recombinants when used with Bacillus subtilis 6GM15, and alpha-complementation with plasmid marker rescue when used with Bacillis subtilis 6GM15[pHPS9R] (ATCC 37818).[3] The cat86::lacZalpha fusion is in-frame. Expression is controlled by the P59 promoter from Lactococcus lactis subsp. cremoris Wg2. [1] This is the preferred strain for isolating the plasmid, because the copy number is higher in E.coli than in B. subtilis. (personal communication) Restriction digests of the clone give the following sizes (kb): BamHI--5.7, EcoRI--5.7, PstI--4.9, 0.7. (ATCC staff) Medium is -1 1065 plus erythromycin (200 ug/ml). Plasmid was not verified by restriction analysis. Strain was checked for the following phenotypes:showed growth on LB + chloramphenicol (5 ug/ml, but not 10 ug/ml); blue colonies on IPTG + Xgal (80 ug/ml); (ATCC staff) blue colonies on IPTG + Xgal (80 ug/ml) + erythromycin (150 ug/ml). (ATCC staff) Medium is -1 1065 plus chloramphenicol (5 ug/ml). pRB374도 ATCC (ATCC® Number: 77374 ™)에서 제공하고 있는 것 같습니다. 참고 논문은 아래와 같습니다. Bruckner R. A series of shuttle vectors for Bacillus subtilis and Eschericia coli. Gene 122: 187-192, 1992. 벡터에 대한 기본설명은 아래와 같습니다. (http://seq.yeastgenome.org/vectordb/vector_descrip/PRB374.html) Deposited by: Reinhold Bruckner Neo confers resistance to neomycin and kanamycin and ble confers resistance to bleomycin and phleomycin. [1] Shuttle expression vector. [1] The Bacillus subtilis promoter vegII initiates transcription in both B. subtilis and E. coli. [1] Structural stability of the plasmid in B. subtilis can be affected by high levels of protein production. Under these conditions, cell growth and stability may be improved by reducing the antibiotic concentration in the media. [1] May not be suitable for cloning very strong expression signals. [1] The order of the major features in the plasmid is: To terminator - vegII promoter - HindIII/MCS/EcoRI - rrnB terminator - ampR - pMB1 ori - pUB110 ori - neoR - bleR. [1] Restriction digests of the clone give the following sizes (kb): EcoRI--5.9; BamHI--5.9; BglI/BglII--3.5, 2.4. (ATCC staff) Medium is 1227 LB plus ampicillin. >Bacillus vector중 pHPS9, pRB374 염기서열이 필요합니다. >자료나. web site좀 알려주세요. >cloning또는 expression 에 관련된 자료도 부탁드립니다.