東華大學圖書館 |

語系: 繁體中文

說明(常見問題)

回圖書館首頁

手機版館藏查詢

登入

回首頁

切換: 標籤 | MARC模式 | ISBD

Identification of immunoreactive pro...

Brouwers, Hubert Johan Marie.

FindBook

Google Book

Amazon

博客來

Identification of immunoreactive protein encoding genes of the fish pathogen Piscirickettsia salmonis and evaluation of their use in genetic vaccination.

紀錄類型:	書目-電子資源 : Monograph/item
正題名/作者:	Identification of immunoreactive protein encoding genes of the fish pathogen Piscirickettsia salmonis and evaluation of their use in genetic vaccination./
作者:	Brouwers, Hubert Johan Marie.
面頁冊數:	187 p.
附註:	Source: Dissertation Abstracts International, Volume: 65-09, Section: B, page: 4346.
Contained By:	Dissertation Abstracts International65-09B.
標題:	Health Sciences, Immunology. -
電子資源:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=NQ93848
ISBN:	0612938484

Identification of immunoreactive protein encoding genes of the fish pathogen Piscirickettsia salmonis and evaluation of their use in genetic vaccination.
Brouwers, Hubert Johan Marie.

Identification of immunoreactive protein encoding genes of the fish pathogen Piscirickettsia salmonis and evaluation of their use in genetic vaccination. - 187 p.

Source: Dissertation Abstracts International, Volume: 65-09, Section: B, page: 4346.

Thesis (Ph.D.)--University of Prince Edward Island (Canada), 2005.

Since the emergence of Piscirickettsia salmonis in the late 1980's, it has become a major disease issue in the salmonid aquaculture and no effective control measures are currently available. As a potential vaccination strategy, genetic or DNA vaccines offer great promise. They function by carrying the gene coding for a protective antigen on a plasmid into the host cells where the antigen is produced by the cells themselves. These vaccines mimic the pathway by which antigens from intracellular pathogens are normally presented to the immune system. To develop a DNA vaccine for P. salmonis , genes coding for potentially protective antigens had to be identified although very little is known about the genetic makeup of this pathogen. The most appropriate way to identify protective genes was by generating an expression library of the entire genome of P. salmonis and screening this library with anti-P. salmonis rabbit antiserum. To obtain large amounts of P. salmonis for genomic DNA extraction and antigen production, the growth in culture of P. salmonis had to be optimized. To follow the course of infection of P. salmonis in CHSE-214 cell culture a sandwich ELISA (s-ELISA) was developed using rabbit anti-P. salmonis antiserum to capture P. salmonis and monoclonal 10E6.6C5 for detection. This s-ELISA proved to be both sensitive and practical. The optimum time to harvest P. salmonis culture was determined to be 10 days post inoculation, with the fastest increase in antigen levels between day 6 and 7. P. salmonis genomic DNA without contaminating CHSE-214 DNA was obtained by treating purified P. salmonis with DNaseI to remove CHSE-214 DNA before P. salmonis lysis and DNA purification. Genomic DNA was used to generate an expression library in the lambdaZapExpress vector. By screening approximately 18,000 clones, 41 immuno-reactive clones were identified. Inserts from these clones were amplified by PCR and sequenced. Ten open reading frames (ORFs) were assigned putative functions by comparing deduced amino acid sequences to sequences available in Genbank. Five clones were selected for showing homologies to promising targets for the immune system. Putative functions of these five ORFs were: (1) the 17 kDa surface antigen from the spotted fever group rickettsia; (2) transposase; (3) ATP binding cassette type transporter; (4) Preprotein translocase subunit SecA; and (5) amino acid transporter. (Abstract shortened by UMI.)

ISBN: 0612938484Subjects--Topical Terms:

1017716
Health Sciences, Immunology.

Identification of immunoreactive protein encoding genes of the fish pathogen Piscirickettsia salmonis and evaluation of their use in genetic vaccination.
LDR:03540nmm 2200313 4500 001 1849804
005 20051203081253.5
008 130614s2005 eng d
020 $a 0612938484
035 $a (UnM)AAINQ93848
035 $a AAINQ93848
040 $a UnM $c UnM
100 1 $a Brouwers, Hubert Johan Marie. $3 1937739
245 1 0 $a Identification of immunoreactive protein encoding genes of the fish pathogen Piscirickettsia salmonis and evaluation of their use in genetic vaccination.
300 $a 187 p.
500 $a Source: Dissertation Abstracts International, Volume: 65-09, Section: B, page: 4346.
500 $a Advisers: R. J. F. Markham; S. R. M. Jones.
502 $a Thesis (Ph.D.)--University of Prince Edward Island (Canada), 2005.
520 $a Since the emergence of Piscirickettsia salmonis in the late 1980's, it has become a major disease issue in the salmonid aquaculture and no effective control measures are currently available. As a potential vaccination strategy, genetic or DNA vaccines offer great promise. They function by carrying the gene coding for a protective antigen on a plasmid into the host cells where the antigen is produced by the cells themselves. These vaccines mimic the pathway by which antigens from intracellular pathogens are normally presented to the immune system. To develop a DNA vaccine for P. salmonis , genes coding for potentially protective antigens had to be identified although very little is known about the genetic makeup of this pathogen. The most appropriate way to identify protective genes was by generating an expression library of the entire genome of P. salmonis and screening this library with anti-P. salmonis rabbit antiserum. To obtain large amounts of P. salmonis for genomic DNA extraction and antigen production, the growth in culture of P. salmonis had to be optimized. To follow the course of infection of P. salmonis in CHSE-214 cell culture a sandwich ELISA (s-ELISA) was developed using rabbit anti-P. salmonis antiserum to capture P. salmonis and monoclonal 10E6.6C5 for detection. This s-ELISA proved to be both sensitive and practical. The optimum time to harvest P. salmonis culture was determined to be 10 days post inoculation, with the fastest increase in antigen levels between day 6 and 7. P. salmonis genomic DNA without contaminating CHSE-214 DNA was obtained by treating purified P. salmonis with DNaseI to remove CHSE-214 DNA before P. salmonis lysis and DNA purification. Genomic DNA was used to generate an expression library in the lambdaZapExpress vector. By screening approximately 18,000 clones, 41 immuno-reactive clones were identified. Inserts from these clones were amplified by PCR and sequenced. Ten open reading frames (ORFs) were assigned putative functions by comparing deduced amino acid sequences to sequences available in Genbank. Five clones were selected for showing homologies to promising targets for the immune system. Putative functions of these five ORFs were: (1) the 17 kDa surface antigen from the spotted fever group rickettsia; (2) transposase; (3) ATP binding cassette type transporter; (4) Preprotein translocase subunit SecA; and (5) amino acid transporter. (Abstract shortened by UMI.)
590 $a School code: 1108.
650 4 $a Health Sciences, Immunology. $3 1017716
650 4 $a Biology, Veterinary Science. $3 1021733
650 4 $a Agriculture, Fisheries and Aquaculture. $3 1020913
650 4 $a Agriculture, Animal Culture and Nutrition. $3 1017857
690 $a 0982
690 $a 0778
690 $a 0792
690 $a 0475
710 2 0 $a University of Prince Edward Island (Canada). $3 1017636
773 0 $t Dissertation Abstracts International $g 65-09B.
790 1 0 $a Markham, R. J. F., $e advisor
790 1 0 $a Jones, S. R. M., $e advisor
790 $a 1108
791 $a Ph.D.
792 $a 2005
856 4 0 $u http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=NQ93848