Abstract：
:Targeted mutagenesis is one of the key methods for functional gene analysis. A simplified variant of gene targeting uses direct microinjection of custom-designed Zinc Finger Nuclease (ZFN) mRNAs into Drosophila embryos. To evaluate the applicability of this method to gene targeting in another insect, we mutagenized the Bombyx mori epidermal color marker gene BmBLOS2, which controls the formation of uric acid granules in the larval epidermis. Our results revealed that ZFN mRNA injection is effective to induce somatic, as well as germline, mutations in a targeted gene by non-homologous end joining (NHEJ). The ZFN-induced NHEJ mutations lack end-filling and blunt ligation products, and include mainly 7 bp or longer deletions, as well as single nucleotide insertions. These observations suggest that the B. mori double-strand break repair system relies on microhomologies rather than on a canonical ligase IV-dependent mechanism. The frequency of germline mutants in G(1) was sufficient to be used for gene targeting relying on a screen based solely on molecular methods.
journal_name
Insect Biochem Mol Bioljournal_title
Insect biochemistry and molecular biologyauthors
Takasu Y,Kobayashi I,Beumer K,Uchino K,Sezutsu H,Sajwan S,Carroll D,Tamura T,Zurovec Mdoi
10.1016/j.ibmb.2010.07.012subject
Has Abstractpub_date
2010-10-01 00:00:00pages
759-65issue
10eissn
0965-1748issn
1879-0240pii
S0965-1748(10)00167-0journal_volume
40pub_type
杂志文章abstract：:A cDNA expression library prepared from mRNA of Haemaphysalis longicornis (H. longicornis) was screened with a H. longicornis-infested rabbit serum. A cDNA encoding 27/30kDa proteins was cloned and designated P27/30 gene. The predicted amino acid sequence of the P27/30 gene shows a rather high homology (58% amino acid...
journal_title：Insect biochemistry and molecular biology
pub_type： 杂志文章
doi：10.1016/s0965-1748(01)00081-9
更新日期：2001-12-01 00:00:00
abstract：:Two bisacylhydrazine insecticides with ecdysone-mimetic action, RH-5849 and RH-5992, have been subjected to several bioassay procedures that are prerequisites for ecdysone action in Drosophila larvae: (a) induction of early ecdysone-specific puffs on the polytene chromosomes of the larval salivary glands; (b) secretio...
journal_title：Insect biochemistry and molecular biology
pub_type： 杂志文章
doi：10.1016/s0965-1748(99)00077-6
更新日期：1999-11-01 00:00:00
abstract：:A cDNA encoding a Na(+)-dependent glutamate transporter has been cloned from the brain of the cockroach Diploptera punctata. The cDNA encodes a transporter protein of 481 amino acids, designated DipEAAT1, which when expressed in baculovirus infected insect cells, resulted in a 40-50 fold increase in [(3)H]L-glutamate ...
journal_title：Insect biochemistry and molecular biology
pub_type： 杂志文章
doi：10.1016/s0965-1748(00)00004-7
更新日期：2000-05-01 00:00:00
abstract：:Phloem-sap feeding Hemipteran insects have access to a sucrose-rich diet but are dependent on sucrose hydrolysis and hexose transport for carbon nutrition. A cDNA library from Nilaparvata lugens (rice brown planthopper) was screened for clones encoding potential transmembrane transporters. A selected cDNA, NlHT1, enco...
journal_title：Insect biochemistry and molecular biology
pub_type： 杂志文章
doi：10.1016/j.ibmb.2007.07.001
更新日期：2007-11-01 00:00:00
abstract：:The importance of Chagas disease motivated the scientific effort to obtain the complete genomic sequence of the vector species Rhodnius prolixus, this information is also relevant to the understanding of triatomine biology in general. The central nervous system is the key regulator of insect physiology and behavior. N...
journal_title：Insect biochemistry and molecular biology
pub_type： 杂志文章
doi：10.1016/j.ibmb.2015.05.003
更新日期：2016-02-01 00:00:00
abstract：:RNA interference is a crucial antiviral mechanism in arthropods, including in mosquito vectors of arthropod-borne viruses (arboviruses). Although the exogenous small interfering RNA (siRNA) pathway constitutes an efficient antiviral response in mosquitoes, virus-derived P-element induced wimpy testis (PIWI)-interactin...
journal_title：Insect biochemistry and molecular biology
pub_type： 杂志文章
doi：10.1016/j.ibmb.2019.04.008
更新日期：2019-06-01 00:00:00
abstract：:The brown planthopper, Nilaparvata lugens, is a model species for hemimetabolous development and the most important pest insect in rice, which is the major staple crop for about half of the world population. Despite its importance, little is known of the N-glycosylation process in this insect. Here we report on the N-...
journal_title：Insect biochemistry and molecular biology
pub_type： 杂志文章
doi：10.1016/j.ibmb.2019.01.008
更新日期：2019-04-01 00:00:00
abstract：:Insects possess a cuticle that covers all tissues exposed to the outside world including the body, the fore- and hindgut and the luminal side of the tracheae. The cuticle is a multifunctional device that protects its carriers against dehydration, arms them against predators, constitutes a physical barrier to prevent p...
journal_title：Insect biochemistry and molecular biology
pub_type： 杂志文章,评审
doi：10.1016/j.ibmb.2010.03.003
更新日期：2010-05-01 00:00:00
abstract：:Phenol oxidase (PO) is known to play an important role in defense mechanisms in insect immunity. It is present as a zymogen in insect hemolymph, and can be activated by a specific proteolytic reaction that is stimulated by microbial cell wall components. The pro-phenol oxidase (pro-PO) purified from the larval hemolym...
journal_title：Insect biochemistry and molecular biology
pub_type： 杂志文章
doi：10.1016/s0965-1748(97)00066-0
更新日期：1997-10-01 00:00:00
abstract：:One promising method to prevent vector-borne diseases is through the use of transmission blocking vaccines (TBVs). However, developing several anti-pathogen TBVs may be impractical. In this study, we have identified a conserved candidate carbohydrate target in the midguts of several Arthropod vectors. A screen of the ...
journal_title：Insect biochemistry and molecular biology
pub_type： 杂志文章
doi：10.1016/j.ibmb.2004.09.005
更新日期：2005-01-01 00:00:00
abstract：:The insect arginine vasopressin-like (AVPL) peptide is of special interest because of its potential function in the regulation of diuresis. Genome sequences of the red flour beetle Tribolium castaneum yielded the genes encoding AVPL and AVPL receptor, whereas the homologous sequences are absent in the genomes of the f...
journal_title：Insect biochemistry and molecular biology
pub_type： 杂志文章
doi：10.1016/j.ibmb.2008.04.006
更新日期：2008-07-01 00:00:00
abstract：:The distributions of mRNAs for two cuticular proteins of Hyalophora cecropia were examined with RT-PCR and in situ hybridization. For major regions of larval and pupal cuticle, there was a strong correspondence between the type of cuticle and the predominant cuticular protein message found. Epidermal cells underlying ...
journal_title：Insect biochemistry and molecular biology
pub_type： 杂志文章
doi：10.1016/j.ibmb.2003.06.010
更新日期：2003-12-01 00:00:00
abstract：:Nicotine is one of the most abundant and toxic secondary plant metabolites in nature and is defined by high toxicity to plant-feeding insects. Studies suggest that increased expression of cytochrome P450 (CYP6CY3) and the homologous CYP6CY4 genes in Myzus persicae nicotianae is correlated with tolerance to nicotine. I...
journal_title：Insect biochemistry and molecular biology
pub_type： 杂志文章
doi：10.1016/j.ibmb.2016.06.002
更新日期：2016-08-01 00:00:00
abstract：:Antifreeze proteins (AFPs) are characterized by their capacity to inhibit the growth of ice and are produced by a variety of polar fish, terrestrial arthropods and other organisms inhabiting cold environments. This capacity reflects their role as stabilizers of supercooled body fluids. The longhorn beetle Rhagium inqu...
journal_title：Insect biochemistry and molecular biology
pub_type： 杂志文章
doi：10.1016/j.ibmb.2010.11.002
更新日期：2011-02-01 00:00:00
abstract：:Two beta-glycosidases (BG) (Mr 47,000 and Mr 50,000) were purified from Spodoptera frugiperda (Lepidoptera: Noctuidae) midguts. These two polypeptides associate or dissociate depending on the medium ionic strength. The Mr 47,000 BG probably has two active sites. One of the putative active sites (cellobiase site) hydro...
journal_title：Insect biochemistry and molecular biology
pub_type： 杂志文章
doi：10.1016/s0965-1748(00)00090-4
更新日期：2000-12-01 00:00:00
abstract：:Using in vitro methods, we investigated the transfer of cholesterol from larval Manduca sexta midgut to the hemolymph lipoprotein, lipophorin, and the transfer of cholesterol from lipophorin to larval fat body. In the midgut, transfer of free cholesterol shows saturation kinetics, but the apparent Km is higher than th...
journal_title：Insect biochemistry and molecular biology
pub_type： 杂志文章
doi：10.1016/s0965-1748(02)00051-6
更新日期：2002-09-01 00:00:00
abstract：:We investigated changes in the pupal transcriptome of the flesh fly Sarcophaga crassipalpis, 3 and 25 h after parasitization by the ectoparasitoid wasp, Nasonia vitripennis. These time points are prior to hatching of the wasp eggs, thus the results document host responses to venom injection, rather than feeding by the...
journal_title：Insect biochemistry and molecular biology
pub_type： 杂志文章
doi：10.1016/j.ibmb.2013.10.003
更新日期：2013-12-01 00:00:00
abstract：:Chitin synthase (EC 2.4.1.16) is a crucial enzyme responsible for chitin biosynthesis in all chitin-containing organisms. This paper reports a complete cDNA encoding chitin synthase 1 (AqCHS1), change of AqCHS1 mRNA level in response to diflubenzuron exposure, and concentration-dependent effect of diflubenzuron on chi...
journal_title：Insect biochemistry and molecular biology
pub_type： 杂志文章
doi：10.1016/j.ibmb.2006.06.002
更新日期：2006-09-01 00:00:00
abstract：:Degenerate oligonucleotides were designed on the basis of conserved amino acid sequences in the DNA binding domains of the ecdysone receptors from Drosophila melanogaster (DmEcR) and Chironomus tentans (CtEcR). Using these oligonucleotides a fragment encoding part of the DNA binding domain of the Lucilia cuprina ecdys...
journal_title：Insect biochemistry and molecular biology
pub_type： 杂志文章
doi：10.1016/s0965-1748(97)00019-2
更新日期：1997-06-01 00:00:00
abstract：:The molt-intermolt cycle is an essential feature in holometabolous and hemimetabolous insects' development. In the silkworm, silk glands are under dramatic morphological and functional changes with fibroin genes' transcription being repeatedly turned off and on during the molt-intermolt cycles. However, the molecular ...
journal_title：Insect biochemistry and molecular biology
pub_type： 杂志文章
doi：10.1016/j.ibmb.2016.07.002
更新日期：2016-09-01 00:00:00
abstract：:Olfaction in Drosophila is mediated by a large family of membrane-bound odorant receptor proteins (Ors). In heterologous cells, we investigated whether the structural features and signalling mechanisms of ligand-binding Drosophila Ors are consistent with them being G protein-coupled receptors (GPCRs). The detailed mem...
journal_title：Insect biochemistry and molecular biology
pub_type： 杂志文章
doi：10.1016/j.ibmb.2008.05.002
更新日期：2008-08-01 00:00:00
abstract：:Bombyx mori, the domesticated silkworm, is a major insect model for research, and the first lepidopteran for which draft genome sequences became available in 2004. Two independent data sets from whole-genome shotgun sequencing were merged and assembled together with newly obtained fosmid- and BAC-end sequences. The re...
journal_title：Insect biochemistry and molecular biology
pub_type： 杂志文章
doi：10.1016/j.ibmb.2008.11.004
更新日期：2008-12-01 00:00:00
abstract：:The midgut of hematophagous insects is the initial site of infection by arthropod-borne viruses (arboviruses) and plays a crucial role in vector competence. To further understand processes that occur in the midgut in response to infection by an arbovirus, DNA microarrays were used to analyze gene expression changes fo...
journal_title：Insect biochemistry and molecular biology
pub_type： 杂志文章
doi：10.1016/j.ibmb.2005.07.006
更新日期：2005-11-01 00:00:00
abstract：:The juvenile hormone analog (JHA) methoprene was used to induce the synthesis of the yolk protein precursor vitellogenin (Vg) in adult females and males of the cockroach Leucophaea maderae. The female- and male-produced vitellogenin (VgF and VgM, respectively) contained polypeptides of 112, 95, 92, and 54 kDa. Also pr...
journal_title：Insect biochemistry and molecular biology
pub_type： 杂志文章
doi：10.1016/s0965-1748(97)00071-4
更新日期：1997-11-01 00:00:00
abstract：:Insect exoskeletons are composed of the cuticle, a biomaterial primarily formed from the linear and relatively rigid polysaccharide, chitin, and structural proteins. This extracellular material serves both as a skin and skeleton, protecting insects from environmental stresses and mechanical damage. Despite its rather ...
journal_title：Insect biochemistry and molecular biology
pub_type： 杂志文章
doi：10.1016/j.ibmb.2017.11.003
更新日期：2017-12-01 00:00:00
abstract：:In addition to its wide role in metabolism, iron in insects has been implicated in vitellogenesis and the immune response. The NRAMP family comprises a well-conserved family of divalent cation transporters in metazoans. To gain insight on the role of NRAMP in Anopheles albimanus, we cloned a cDNA encoding a 571-residu...
journal_title：Insect biochemistry and molecular biology
pub_type： 杂志文章
doi：10.1016/j.ibmb.2007.02.009
更新日期：2007-06-01 00:00:00
abstract：:To gain insight into the molecular repertoire of the adult female salivary glands of the tiger mosquito Aedes albopictus, we performed transcriptome and proteome analysis. cDNA clones were sequenced and assembled in clusters of related sequences and the corresponding genes assigned to one of three categories: housekee...
journal_title：Insect biochemistry and molecular biology
pub_type： 杂志文章
doi：10.1016/j.ibmb.2006.10.007
更新日期：2007-02-01 00:00:00
abstract：:To analyze cecropin B promoter (P-CecB) activity in vivo, we constructed transgenic silkworms that expressed EGFP under the control of P-CecB using the piggyBac transposable element. Genomic Southern blot analysis of the G1 and G2 generations indicated the stable insertion of EGFP in the genome. Injection of Escherich...
journal_title：Insect biochemistry and molecular biology
pub_type： 杂志文章
doi：10.1016/j.ibmb.2006.03.002
更新日期：2006-05-01 00:00:00
abstract：:Juvenile hormone (JH) titers must be modulated to permit the normal progress of development and reproduction in mosquitoes. In adult female Aedes aegypti, JH levels are low at adult eclosion, elevated in sugar-fed females and low again after a blood meal. Although degradation plays a role, JH titer is fundamentally de...
journal_title：Insect biochemistry and molecular biology
pub_type： 杂志文章,评审
doi：10.1016/j.ibmb.2004.03.021
更新日期：2004-07-01 00:00:00
abstract：:A cDNA clone containing a 921 bp open-reading frame (307 amino acids; 34 kDa) homologous to the TATA-binding protein (TBP) was isolated and sequenced from a Spodoptera frugiperda cell line that is commonly used in the baculovirus expression system. Analysis of the S. frugiperda TBP (SfTBP) sequence showed that the ami...
journal_title：Insect biochemistry and molecular biology
pub_type： 杂志文章
doi：10.1016/0965-1748(94)90057-4
更新日期：1994-07-01 00:00:00
