Insert Size (insert + size)

Distribution by Scientific Domains


Selected Abstracts


Construction of a cDNA library of Bemisia tabaci for use in the ,yeast two-hybrid screen' method

EPPO BULLETIN, Issue 1 2002
S. Ohnesorge
The molecular mechanisms involved in the circulative, non-propagative transmission pathway of TYLCV through its vector the whitefly Bemisia tabaci have hardly been studied. Points requiring investigation include the specific adhesion of virus coat protein to insect structures, the proteins involved in membrane passage in the insect and the possibility of replication of the virus in the vector. To isolate the insect proteins which are involved in transmission by interaction with viral proteins, we propose to use the ,yeast two-hybrid screen' genetic method. For this method, it is indispensable to have a ,cDNA library' of the organism concerned, cloned in plasmids, and our first step has been to develop this. A new method was developed for isolating whitefly mRNA. From this mRNA, cDNA was synthesized, ligated in the plasmid pGADT7 (Clontech) and transformed in bacteria to amplify the plasmid DNA. The number of independent clones and average insert size of the plasmids were determined. [source]


Artificial chromosome libraries of Streptomyces coelicolor A3(2) and Planobispora rosea,

FEMS MICROBIOLOGY LETTERS, Issue 1 2003
Rosa Alduina
Abstract Using an Escherichia coli,Streptomyces shuttle vector derived from a bacterial artificial chromosome (BAC), we developed methodologies for the construction of BAC libraries of filamentous actinomycetes. Libraries of Streptomyces coelicolor, the model actinomycete, and Planobispora rosea, a genetically intractable strain, were constructed. Both libraries have an average insert size of 60 kb, with maximal insert larger than 150 kb. The S. coelicolor library was evaluated by selected hybridisations to Dra I fragments and by end sequencing of a few clones. Hybridisation of the P. rosea library to selected probes indicates a good representation of the P. rosea genome and that the library can be used to facilitate the genomic analysis of this actinomycete. [source]


Effects of Processing Conditions on Qualities of Rice Fries

JOURNAL OF FOOD SCIENCE, Issue 4 2001
R.S. Kadan
ABSTRACT Two rice flour mixtures, 2 extruding temperatures, and 2 insert sizes were used to develop rice fries. The extruded material was cut into pieces 7 to 8-cm long, prefried in rice oil at 180°C for 20 s, and kept frozen until final frying for another 70 s. The rice fries were evaluated 5 min and 10 min after final frying for their lipid, moisture, and instrumental texture characteristics. Extruding temperature, rice cultivar, and insert size were significantly related to fat content, moisture, hardness, and fracturability values. A mixture consisting of 80:20 (waxy:long-grain), and extruded at 70 °C using a 6 mm insert, 5 min after frying, gave a texture profile analysis value for hardness, cohesive-ness, and gumminess values comparable to commercial potato French fries. The rice fries made by the process also had less than 50% fat than potato fries. [source]


Efficient cloning of plant genomes into bacterial artificial chromosome (BAC) libraries with larger and more uniform insert size

PLANT BIOTECHNOLOGY JOURNAL, Issue 3 2004
Boulos Chalhoub
Summary The construction of bacterial artificial chromosome (BAC) libraries remains relatively complex and laborious, such that any technological improvement is considered to be highly advantageous. In this study, we addressed several aspects that improved the quality and efficiency of cloning of plant genomes into BACs. We set the ,single tube vector' preparation method with no precipitation or gel electrophoresis steps, which resulted in less vector DNA damage and a remarkable two- to threefold higher transformation efficiency compared with other known vector preparation methods. We used a reduced amount of DNA for partial digestion (up to 5 µg), which resulted in less BAC clones with small inserts. We performed electrophoresis in 0.25 × TBE (Tris, boric acid, ethylenediaminetetraacetic acid) buffer instead of 0.5 × TBE, which resulted in larger and more uniformly sized BAC inserts and, surprisingly, a two- to threefold higher transformation efficiency, probably due to less contamination with borate ions. We adopted a triple size selection that resulted in an increased mean insert size of up to 70 kb and a transformation efficiency comparable with that of double size selection. Overall, the improved protocol presented in this study resulted in a five- to sixfold higher cloning efficiency and larger and more uniformly sized BAC inserts. BAC libraries with the desired mean insert size (up to 200 kb) were constructed from several plant species, including hexaploid wheat. The improved protocol will render the construction of BAC libraries more available in plants and will greatly enhance genome analysis, gene mapping and cloning. [source]


Effects of Processing Conditions on Qualities of Rice Fries

JOURNAL OF FOOD SCIENCE, Issue 4 2001
R.S. Kadan
ABSTRACT Two rice flour mixtures, 2 extruding temperatures, and 2 insert sizes were used to develop rice fries. The extruded material was cut into pieces 7 to 8-cm long, prefried in rice oil at 180°C for 20 s, and kept frozen until final frying for another 70 s. The rice fries were evaluated 5 min and 10 min after final frying for their lipid, moisture, and instrumental texture characteristics. Extruding temperature, rice cultivar, and insert size were significantly related to fat content, moisture, hardness, and fracturability values. A mixture consisting of 80:20 (waxy:long-grain), and extruded at 70 °C using a 6 mm insert, 5 min after frying, gave a texture profile analysis value for hardness, cohesive-ness, and gumminess values comparable to commercial potato French fries. The rice fries made by the process also had less than 50% fat than potato fries. [source]