Building Scarless Gene Libraries in the Chromosome of Bacteria

Author(s):  
Gol Mohammad Dorrazehi ◽  
Sebastian Worms ◽  
Jason Baby Chirakadavil ◽  
Johann Mignolet ◽  
Pascal Hols ◽  
...  
Keyword(s):  
2000 ◽  
Vol 28 (5) ◽  
pp. A365-A365
Author(s):  
S. A. Richmond ◽  
A. D. Trowbridge ◽  
M. Reid
Keyword(s):  

1984 ◽  
Vol 4 (10) ◽  
pp. 1961-1969
Author(s):  
J Leavitt ◽  
P Gunning ◽  
P Porreca ◽  
S Y Ng ◽  
C S Lin ◽  
...  

There are more than 20 beta-actin-specific sequences in the human genome, many of which are pseudogenes. To facilitate the isolation of potentially functional beta-actin genes, we used the new method of B. Seed (Nucleic Acids Res. 11:2427-2446, 1983) for selecting genomic clones by homologous recombination. A derivative of the pi VX miniplasmid, pi AN7 beta 1, was constructed by insertion of the 600-base-pair 3' untranslated region of the beta-actin mRNA expressed in human fibroblasts. Five clones containing beta-actin sequences were selected from an amplified human fetal gene library by homologous recombination between library phage and the miniplasmid. One of these clones contained a complete beta-actin gene with a coding sequence identical to that determined for the mRNA of human fibroblasts. A DNA fragment consisting of mostly intervening sequences from this gene was then used to identify 13 independent recombinant copies of the analogous gene from two specially constructed gene libraries, each containing one of the two types of mutant beta-actin genes found in a line of neoplastic human fibroblasts. The amino acid and nucleotide sequences encoded by the unmutated gene predict that a guanine-to-adenine transition is responsible for the glycine-to-aspartic acid mutation at codon 244 and would also result in the loss of a HaeIII site. Detection of this HaeIII polymorphism among the fibroblast-derived clones verified the identity of the beta-actin gene expressed in human fibroblasts.


2010 ◽  
Vol 76 (6) ◽  
pp. 1740-1745 ◽  
Author(s):  
Ming Tang ◽  
Lu Lv ◽  
Shengli Jing ◽  
Lili Zhu ◽  
Guangcun He

ABSTRACT The brown planthopper (Nilaparvata lugens Stål), the most destructive pest of rice, has been identified, including biotypes with high virulence towards previously resistant rice varieties. There have also been many reports of a yeast-like symbiont of N. lugens, but little is known about the bacterial microbes. In this study, we examined the bacterial microbes in N. lugens and identified a total of 18 operational taxonomic units (OTUs) representing four phyla (Proteobacteria, Firmicutes, Actinobacteria, and Bacteroidetes) by sequencing and analyzing 16S rRNA gene libraries obtained from three populations of N. lugens, which were maintained on the rice varieties TN1, Mudgo, and ASD7. Several of the OTUs were similar to previously reported secondary symbionts of other insects, including an endosymbiont of the psyllid Glycapsis brimblecombei, an Asaia sp. found in the mosquito Anopheles stephensi, and Wolbachia, found in the mite Metaseiulus occidentalis. However, the species and numbers of the detected OTUs differed substantially among the N. lugens populations. Further, in situ hybridization analysis using digoxigenin-labeled probes indicated that OTU 1 was located in hypogastrium tissues near the ovipositor and ovary in biotype 1 insects, while OTU 2 was located in the front of the ovipositor sheath in biotype 2 insects. In addition, masses of bacterium-like organisms were observed in the tubes of salivary sheaths in rice plant tissues that the insects had fed upon. The results provide indications of the diversity of the bacterial microbes harbored by the brown planthopper and of possible associations between specific bacterial microbes and biotypes of N. lugens.


1987 ◽  
Vol 3 (4) ◽  
pp. 120-121 ◽  
Author(s):  
L. Grodzinski
Keyword(s):  

1987 ◽  
Vol 43 (1) ◽  
pp. 53-56 ◽  
Author(s):  
Margaret E. Collins ◽  
Michael T. Moss ◽  
Sue Wall ◽  
Jeremy W. Dale
Keyword(s):  

BioTechniques ◽  
2018 ◽  
Vol 64 (1) ◽  
Author(s):  
Bettina Rohweder ◽  
Florian Semmelmann ◽  
Christiane Endres ◽  
Reinhard Sterner

Sign in / Sign up

Export Citation Format

Share Document