Cloning and expression of the Legionella micdadei "common antigen" in Escherichia coli
Research output: Contribution to journal › Journal article › Research › peer-review
Standard
Cloning and expression of the Legionella micdadei "common antigen" in Escherichia coli. / Bangsborg, Jette Marie; Collins, M T; Høiby, N; Hindersson, P.
In: A P M I S. Acta Pathologica, Microbiologica et Immunologica Scandinavica, Vol. 97, No. 1, 1989, p. 14-22.Research output: Contribution to journal › Journal article › Research › peer-review
Harvard
APA
Vancouver
Author
Bibtex
}
RIS
TY - JOUR
T1 - Cloning and expression of the Legionella micdadei "common antigen" in Escherichia coli
AU - Bangsborg, Jette Marie
AU - Collins, M T
AU - Høiby, N
AU - Hindersson, P
PY - 1989
Y1 - 1989
N2 - To study individual Legionella antigens, a Legionella micdadei genomic library in Escherichia coli SC181 was established. Partially Sau3A digested L. micdadei DNA fragments (15-25 kilobase pairs (kb] were cloned into the tetracycline resistance gene of the cosmid vector pHC79. Four thousand ampicillin resistant recombinants were obtained; seven hundred were screened for expression of Legionella antigens in Western blot analysis with a polyspecific E. coli-absorbed anti-L. micdadei rabbit antibody. One of the positive clones expressed a 60 kilodalton (K) antigen, which reacted strongly with a monospecific rabbit antiserum raised against L. micdadei "common antigen" (CA), and an additional 13 K L. micdadei protein. The region encoding these two proteins from the 17 kb recombinant plasmid (pBA 2) was subcloned in pBGS18+. The DNA sequence of the CA encoding region in the 2.7 kb subcloned fragment will provide important information with respect to genetic vs. antigenic relatedness among Legionellae and other Gram-negative species, as well as to CA structure and possible function.
AB - To study individual Legionella antigens, a Legionella micdadei genomic library in Escherichia coli SC181 was established. Partially Sau3A digested L. micdadei DNA fragments (15-25 kilobase pairs (kb] were cloned into the tetracycline resistance gene of the cosmid vector pHC79. Four thousand ampicillin resistant recombinants were obtained; seven hundred were screened for expression of Legionella antigens in Western blot analysis with a polyspecific E. coli-absorbed anti-L. micdadei rabbit antibody. One of the positive clones expressed a 60 kilodalton (K) antigen, which reacted strongly with a monospecific rabbit antiserum raised against L. micdadei "common antigen" (CA), and an additional 13 K L. micdadei protein. The region encoding these two proteins from the 17 kb recombinant plasmid (pBA 2) was subcloned in pBGS18+. The DNA sequence of the CA encoding region in the 2.7 kb subcloned fragment will provide important information with respect to genetic vs. antigenic relatedness among Legionellae and other Gram-negative species, as well as to CA structure and possible function.
KW - Antigens, Bacterial
KW - Bacterial Proteins
KW - Cloning, Molecular
KW - Escherichia coli
KW - Immunoelectrophoresis, Two-Dimensional
KW - Legionella
KW - Molecular Weight
KW - Restriction Mapping
M3 - Journal article
C2 - 2643977
VL - 97
SP - 14
EP - 22
JO - A P M I S. Acta Pathologica, Microbiologica et Immunologica Scandinavica
JF - A P M I S. Acta Pathologica, Microbiologica et Immunologica Scandinavica
SN - 0903-4641
IS - 1
ER -
ID: 40334435