实验基础

Detection of Helicobacter pylori in specimens obtained by endoscopy requires the gastroenterologist to select suitable patients for endoscopy, and to take an adequate number of biopsy specimens; there must also be correct cleaning of the biopsy forceps.

A number of methods are currently available for the detection of Helicobacter pylori, including serology, culture, histology, and isotope breath tests. All have relative advantages and disadvantages of sensitivity, specificity, convenience, expense, and immediacy. The polyme ...

Helicobacter pylori is a curved Gram-negative bacillus that infects the human gastric mucosa. Most methods used to diagnose H. pylori require the collection of gastric samples by biopsy during gastroduodenoscopy. A positive culture or visualization of the organism by histologic exa ...

A crucial step in the baculovirus expression system technique is the selection of recombinant viruses. In most cases positive clones, i.e., recombinant viruses, are detected via the presence of foreign DNA, or by exploiting the phenotypic differences between wild-type viral plaques, wh ...

Protocols for the isolation of recombinant baculoviruses and their use for foreign gene expression in insect cells have been described in detail in the preceding chapters (also see refs. 1–5). Generally, this approach involves the replacement of viral DNA sequences encoding the polyhed ...

Stirred-tank fermentors were originally designed to perform microbial cultures (1). When the need arose for adequate, posttranslationally modified proteins, molecular biologists and bioprocess engineers turned to mammalian and insect cell culture using these fermentors ...

The recombinant baculovirus expression system, developed in the laboratories of Summers (1) and Miller (2), has been widely used for the production of heterologous proteins (3, 4). In this system, the gene to be expressed is cloned into a plasmid transfer vector, downstream of a strong baculovir ...

Baculoviruses have been isolated from a wide range of invertebrates. Their development as pest control agents spans over 40 years, although records of their discovery and use date from considerably earlier (1). The baculoviruses from Lepidoptera (butterflies and moths), Hymenoptera ...

A large number of eukaryotic proteins have been successfully overproduced in insect cells infected with a natural pathogen, a baculovirus, in which foreign gene coding sequences were placed under the control of the viral polyhedrin gene promoter. This expression system is effective be ...

Baculovirus expression vector systems based on the nuclear polyhedrosis viruses of Autographa californica (AcNPV) and Bombyx mori (BmNPV) are in wide use (1). Our laboratory originally designed the BmNPV system with the objective of using the silkworm, B. mori, as an in vivo host (1a, 2). The silkw ...

The acquired immune deficiency syndrome (AIDS) is caused by human immunodeficiency virus type 1 or 2 (HIV-l or HIV-2). The disease is characterized by a high susceptibility to opportunistic infections or to malignant diseases, such as Kaposi’s sarcoma. The major immunologic abnormality is ...

Baculovirus vectors have been developed for the expression of foreign genes in insect cells. Protein yields are often significantly higher than in bacterial, yeast, or vertebrate expression systems (12). It was therefore of interest to test the capacity of this system to direct the complex a ...

The baculovirus expression system represents an elegant means whereby high-level expression of foreign genes can be achieved using eukaryotic insect cells (1,2). Viruses from the Baculoviridae subfamily of insect viruses, usually Autographa californica nuclear polyhedro ...

Over the last 10 years, baculovirus expression vectors have become a very popular and effective means with which to produce recombinant proteins in large quantities (1–5). Posttranslational modifications of the gene products of these insect viruses closely parallel glycosylatio ...

The binding of ligand to the extracellular domain of the epidermal growth factor (EGF) receptor is the event that initiates the activation of the tyrosine kinase activity present on the cytoplasmic domain. This activation leads to the phosphorylation of the receptor and other cellular sub ...

Myelin-associated glycoprotein (MAG) was first identified as the major glycoprotein in the central nervous system (CNS) through 3Hfucose-labeling experiments (1). It is expressed on the surface of glial cells of both the CNS and peripheral nervous system (PNS). During development, it is ...

Almost 50 years ago, Grace reported the successful establishment of continuous cell lines from insects that were without defined biological and biophysical parameters (1 ). Since that time, the minimal distinctive parameters for the establishment, maintenance, and manipulation ...

Ever since the early reports of Goldschmidt (1) and Trager (2) on the in vitro cultivation of lepidopteran tissues, researchers have been interested in using lepidopteran cell cultures for investigations in insect pathology, genetics, and physiology. With the successful development ...

The levels of many recombinant proteins produced using the baculovirus expression vector (BEV) system are extraordinarily high (1, 2), in many cases approaching 1000 mg/L, the level of polyhedrin expression. Unfortunately, other proteins that have been expressed in the BEV system are pro ...

Historically, insect cell culture was considered a method for the production of viral pesticides (1, 2). To support basic research and to explore potential replication of entomopathogenic viruses in vitro, there have been over 400 cell lines established to date from various insect species ...