Alexander N. Glazer

Bio: Alexander N. Glazer is an academic researcher from University of California, Berkeley. The author has contributed to research in topics: Phycobilisome & Phycocyanin. The author has an hindex of 71, co-authored 208 publications receiving 21068 citations. Previous affiliations of Alexander N. Glazer include Pasteur Institute & University of California, Los Angeles.

Cassette Labeling for Facile Construction of Energy Transfer Fluorescent Primers

Abstract: DNA primer sets, labeled with two fluorescent dyes to exploit fluorescence energy transfer (ET), can be efficiently excited with a single laser line and emit strong fluorescence at distinctive wavelengths. Such ET primers are superior to single fluorophore-labeled primers for DNA sequencing and other multiple color-based analyses [J. Ju, C. Ruan, C. W. Fuller, A. N. Glazer and R. A. Mathies (1995) Proc. Natl. Acad. Sci. USA 92, 4347-4351]. We describe here a novel method of constructing fluorescent primers using a universal ET cassette that can be incorporated by conventional synthesis at the 5'-end of an oligonucleotide primer of any sequence. In this cassette, the donor and acceptor fluorophores are separated by a polymer spacer (S6) formed by six 1',2'-dideoxyribose phosphate monomers (S). The donor is attached to the 5' side of the ribose spacer and the acceptor to a modified thymidine attached to the 3' end of the ribose spacer in the ET cassette. The resulting primers, labeled with 6-carboxy-fluorescein as the donor and other fluorescein and rhodamine dyes as acceptors, display well-separated acceptor emission spectra with 2-12-fold enhanced fluorescence intensity relative to that of the corresponding single dye-labeled primers. With single- stranded M13mp18DNA as the template, a typical run with these ET primers on a capillary sequencer provides DNA sequences with 99% accuracy in the first 550 bases using the same amount of DNA template as that typically required using a four-color slab gel automated sequencer.

An Unusual Phycoerythrin from a Marine Cyanobacterium

Abstract: Phycoerythrin conjugates are reagents for cell sorting and analyses in which the argon-ion laser line at 488 nanometers is used for excitation. Many marine Synechococcus strains contain phycoerythrins with absorption maxima at approximately 490 and 550 nanometers; these maxima indicate the presence of phycourobilin and phycoerythrobilin prosthetic groups in the protein. Phycoerythrins of red algae contain both groups, but those of freshwater and soil cyanobacteria contain only phycoerythrobilin. Phycoerythrin purified from Synechococcus WH8103 has molecular properties typical of red algal phycoerythrins, but its phycourobilin content is higher than that of other phycoerythrins. The protein has absorption maxima at 492 and 543 nanometers and corresponding molar extinction coefficients of 2.78 and 1.14 x 106; it fluoresces maximally at 565 nanometers with a quantum yield of 0.5. Conjugates of Synechococcus WH8103 phycoerythrin could increase the sensitivity of cell analysis techniques to almost twice that possible with other phycoerythrin conjugates.

Heterodimeric DNA-binding dyes designed for energy transfer: synthesis and spectroscopic properties

Abstract: Heterodimeric dyes are described which bind tightly to double-stranded (dsDNA) with large fluorescence enhancements. These dyes are designed to exploit energy transfer between donor and acceptor chromophores to tune the separation between excitation and emission wavelengths. The dyes described here absorb strongly at the 488 nm argon ion line, but emit at different wavelengths, and can be applied to multiplex detection of various targets. The chromophores in these dyes, a thiazole orange-thiazole blue heterodimer (TOTAB), two different thiazole orange-ethidium heterodimers (TOED1 and TOED2), and a fluorescein-ethidium heterodimer (FED), are in each case linked through polymethyleneamine linkers. The emission maxima of the DNA-bound dyes lie at 662 (TOTAB), 614 (TOED 2), and 610 nm (FED). The dyes showed a > 100 fold enhancement of the acceptor chromophore fluorescence on binding to dsDNA and no sequence selectivity. In comparison with direct 488 nm excitation of the constituent monomeric dyes, in the heterodimers the fluorescence of the acceptor chromophores was greatly enhanced and the emission of the donor chromophores quenched by over 90%. The acceptor emission per DNA-bound dye molecule was constant from 100 DNA bp:dye to 20 bp:dye and decreased sharply at higher dye:DNA ratios.

14 Papain and Other Plant Sulfhydryl Proteolytic Enzymes

Abstract: Publisher Summary Papain is a simple protein containing only amino acids and devoid of carbohydrate All of the usual amino acids are present with the exception of methionine Papain contains no chromophoric groups other than its constituent amino acids It is rich in tyrosine and tryptophan The amount of active enzyme in papain solutions can be determined by a number of methods Finkle and Smith showed that the sulfhydryl (SH) titer of papain was a direct measure of the amount of active enzyme present Papain is routinely assayed in the presence of freshly prepared 0005 M cysteine and 0001 M ethylenediaminetetraacetic acid (EDTA) The mechanism of action of papain, ficin, chymopapain, and bromelain is very similar While the sequences near the essential thiol groups in these enzymes display varying degrees of homology, judgment as to whether all of these enzymes arose from a common evolutionary precursor has to await more extensive information on their amino acid sequences These enzymes are all activated by SH compounds and cyanide and inactivated by mild oxidizing agents

Initial sequencing and analysis of the human genome.

Abstract: The human genome holds an extraordinary trove of information about human development, physiology, medicine and evolution. Here we report the results of an international collaboration to produce and make freely available a draft sequence of the human genome. We also present an initial analysis of the data, describing some of the insights that can be gleaned from the sequence.

疟原虫var基因转换速率变化导致抗原变异[英]／Paul H, Robert P, Christodoulou Z, et al//Proc Natl Acad Sci U S A

Abstract: 抗原变异可使得多种致病微生物易于逃避宿主免疫应答。表达在感染红细胞表面的恶性疟原虫红细胞表面蛋白1（PfPMP1）与感染红细胞、内皮细胞、树突状细胞以及胎盘的单个或多个受体作用，在黏附及免疫逃避中起关键的作用。每个单倍体基因组var基因家族编码约60种成员，通过启动转录不同的var基因变异体为抗原变异提供了分子基础。

Semiconductor Nanocrystals as Fluorescent Biological Labels

Abstract: Semiconductor nanocrystals were prepared for use as fluorescent probes in biological staining and diagnostics. Compared with conventional fluorophores, the nanocrystals have a narrow, tunable, symmetric emission spectrum and are photochemically stable. The advantages of the broad, continuous excitation spectrum were demonstrated in a dual-emission, single-excitation labeling experiment on mouse fibroblasts. These nanocrystal probes are thus complementary and in some cases may be superior to existing fluorophores.

Quantum Dot Bioconjugates for Ultrasensitive Nonisotopic Detection

Abstract: Highly luminescent semiconductor quantum dots (zinc sulfide-capped cadmium selenide) have been covalently coupled to biomolecules for use in ultrasensitive biological detection. In comparison with organic dyes such as rhodamine, this class of luminescent labels is 20 times as bright, 100 times as stable against photobleaching, and one-third as wide in spectral linewidth. These nanometer-sized conjugates are water-soluble and biocompatible. Quantum dots that were labeled with the protein transferrin underwent receptor-mediated endocytosis in cultured HeLa cells, and those dots that were labeled with immunomolecules recognized specific antibodies or antigens.