geoscience.net logo
+ Resolve Article
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter

+ Translate
+ Subscribe to Site Feed
GeoScience Most Shared ContentMost Shared Content

Overproduction, preparation of monoclonal antibodies and purification of E. coli asparagine synthetase A


, : Overproduction, preparation of monoclonal antibodies and purification of E. coli asparagine synthetase A. Protein Engineering 5(3): 279-283

In order to explore the structure--function relationship of the Escherichia coli asparagine synthetase A it was necessary to devise a system for overexpression of the gene and purification of the gene product. The E. coli asparagine synthetase A structural gene was fused to the 3' end of the human carbonic anhydrase II structural gene and overexpressed in E. coli. The gene product, a 66 kDa fusion protein, which exhibited asparagine synthetase activity, was purified in a single step by affinity chromatography and used as the antigen for the production of monoclonal antibodies. The monoclonal antibodies were screened by ELISA. Colonies were chosen which were positive for purified fusion protein and negative for purified human carbonic anhydrase II. The E. coli asparagine synthetase A gene was then overexpressed and the gene product was used without purification for the final screen. The antibodies selected were used for immunoaffinity chromatography to purify the recombinant overexpressed E. coli asparagine synthetase A. Thus, a procedure is now available so that asparagine synthetase A can be purified to homogeneity in a single step.

US$29.90

PMID: 1357656


Other references

Leiner, G.C.; Abramowitz, S.; Small, M.J., 1970: Arterial blood pH. Review of Allergy 24(5): 439-440

Quintero M.; Chatellier J.Y.; Dunn T.; Flores G., 1997: Quartz overgrowth in Lake Maracaibo, the questionable reliability of petrography revealed by cathodoluminescene analysis. Pages 191-199 1997

Kawakubo, A.; Ariyoshi, S.; Fukui, S.; Shimada, M.; Haseba, S.; Gotoh, Y., 1989: Anesthetic management of a patient with acromegaly complicated with hyperthyroidism. A rare anesthetic experience of a 30-year-old woman with acromegaly complicated with Basedow's disease is reported. After the thyroid function was successfully controlled by drug therapy, resection of pituitary adenoma was performed under gen...

Kotrla M.B.; Murray H.D., 1980: Correlations of shell measurements sex and trematode infection in tarebia granifera in texas usa. Bulletin of the American Malacological Union Incorporated 46: 70

Butler, J.A.; Colles, C.M.; Dyson, S.J.; Kold, S.E.; Poulos, P.W., 2000: Clinical radiology of the horse. Despite the title, this is about radiography of the equine skeleton and, to a lesser extent, internal organs. The new edition contains much additional material, extra illustrations and recent references to the literature (accumulated since 1993).

Göksu, H.Y.; Semiochkina, N.; Shiskina, E.A.; Wieser, A.; El-Faramawy, N.A.; Degteva, M.O.; Jacob, P.; Ivanov, D.V., 2002: Thin layer alpha-Al2O3:C beta dosemeters for the assessment of current dose rate in teeth due to 90Sr intake, and comparison with electron paramagnetic resonance dosimetry. The use of thin-layer alpha-Al2O3:C thermoluminescent detectors (TLDs) for the assessment of current beta dose rate in human teeth due to 90Sr intake is investigated. The teeth used in this study were collected from members of the Techa river popu...

Herber R.; Nuppenau V., 1985: Deep-sea ocean bottom seismographs. Jahrestagung der Deutschen Geophysikalischen Gesellschaft eV

Ioanidi, I.P., 1963: Spring wheat in the Orenburg province.. This collection of papers on wheat cultivation in the above region contains information on local varieties, including data on the chemical and technological properties of their grain.

Huang, L-Min.; Lin, T-Yien.; Chiu, C-Hsun.; Chiu, N-Chang.; Chen, P-Yen.; Yeh, S-Jen.; Boaz, M.; Hutagalung, Y.; Bouckenooghe, A.; Feroldi, E., 2015: Concomitant administration of live attenuated Japanese encephalitis chimeric virus vaccine (JE-CV) and measles, mumps, rubella (MMR) vaccine: randomized study in toddlers in Taiwan. Japanese encephalitis (JE) is the most important cause of viral encephalitis in Asia. In this randomized, open-label, multicenter trial in 550 children aged 12 to 18 months in Taiwan, children received one dose of JE-CV and one dose of MMR vaccine...

Velarde, J.J.; Varney, K.M.; Inman, K.G.; Farfan, M.; Dudley, E.; Fletcher, J.; Weber, D.J.; Nataro, J.P., 2007: Solution structure of the novel dispersin protein of enteroaggregative Escherichia coli. Enteroaggregative Escherichia coli (EAEC), increasingly recognized as an important cause of infant and travelers' diarrhoea, exhibits an aggregative, stacked-brick pattern of adherence to epithelial cells. Adherence is mediated by aggregative...