共查询到20条相似文献,搜索用时 31 毫秒
1.
Zilong Wang Xingfu Zha Ningjia He Zhonghuai Xiang Qingyou Xia 《Molecular biology reports》2010,37(5):2525-2531
RBP1 is an important splicing factor involved in alternative splicing of the pre-mRNA of Drosophila sex-determining gene dsx. In this work, the Bombyx mori homologue of the rbp1 gene, Bmrbp1, was cloned. The pre-mRNA of Bmrbp1 gene is alternatively spliced to produce four mature mRNAs, named Bmrbp1-PA, Bmrbp1-PB, Bmrbp1-PC and Bmrbp1-PD, with nucleotide lengths of 799 nt, 1,316 nt, 894 nt and 724 nt, coding for 142 aa, 159 aa, 91 aa and 117 aa, respectively.
BmRBP1-PA and BmRBP1-PD contain a N terminal RNA recognization motif (RRM) and a C terminal arginine/serine-rich domain, while
BmRBP1-PB and BmRBP1-PC only share a RRM. Amino acid sequence alignments showed that BmRBP1 is conserved with its homologues
in other insects and with other SR family proteins. The RT-PCR showed that Bmrbp1-PA was strongly expressed in all examined tissues and development stages, but Bmrbp1-PB was weakly expressed in these tissues and stages. The expression of both Bmrbp1-PA and Bmrbp1-PB showed no obvious sex difference. While the Bmrbp1-PC and Bmrbp1-PD were beyond detection by RT-PCR very likely due to their tissue/stage specificity. These results suggested that Bmrbp1 should be a member of SR family splicing factors, whether it is involved in the sex-specific splicing of Bmdsx pre-mRNA needs further research. 相似文献
2.
Endoplasmic Reticulum Stress Response of <Emphasis Type="Italic">Bombyx Mori</Emphasis> Calreticulin
Goo TW Park S Jin BR Yun EY Kim I Nho SK Kang SW Kwon OY 《Molecular biology reports》2005,32(3):133-139
We isolated a calreticulin cDNA from the silkworm, Bombyx mori. The cDNA encodes 398 amino acid residues of B. mori calreticulin, with an endoplasmic reticulum retentional HDEL motif at its C-terminus and a predicted molecular mass of 45,801 Da. The B. mori calreticulin shows high protein homology with calreticulin from G. mellonella (88%), A. aegypti (71%), D. melanogaster (69%) and H. sapiens (63%). The highest level of mRNA expression of B. mori calreticulin was exhibited in the fat body of this insect. Although expression of B. mori calreticulin was affected by disturbances in intracellular calcium levels, other ER stress conditions such as inhibition of intracellular protein transport, reduction of disulfide formation, glycosylation inhibition, heat shock and oxidative stress did not disrupt induction of B. mori calreticulin. 相似文献
3.
Junwen Ai Quanyou Yu Tingcai Cheng Fangyin Dai Xuesong Zhang Yong Zhu Zhonghuai Xiang 《Molecular biology reports》2010,37(3):1657-1664
Based on the advances in the silkworm genome project, a new genome-wide analysis of cytochrome P450 genes was performed, focusing
mainly on gene duplication. All four CYP9A subfamily members from the silkworm, Bombyx mori, were cloned by RT-PCR and designated CYP9A19–CYP9A22 by the P450 Nomenclature Committee. They each contain an open reading frame of 1,593 bp in length and encode a putative polypeptide
of 531 amino acids. Both nucleic acid and amino acid sequences share very high identities with one another. The typical motifs
of insect cytochrome P450, including the heme-binding region, helix-C, helix-I, helix-K, and PERF, show high sequence conservation
among the multiple proteins. Alignment with their cDNA sequences revealed that these paralogues share identical gene structures,
each comprising ten exons and nine introns of variable sizes. The locations of their introns (all nine introns follow the
GT–AG rule) are absolutely conserved. CYP9A19, CYP9A20, and CYP9A21 form a tandem cluster on chromosome 17, whereas CYP9A22 is separated from the cluster by four tandem alcohol-dehydrogenase-like genes. Their phylogenetic relationships and structural
comparisons indicated that these paralogues arose as the results of gene duplication events. RT-PCR detected their mRNAs in
different “first line of defense” tissues, as well as in several other organs, suggesting diverse functions. Tissue-selective
expression also indicates their functional divergence. The identified CYP9A genes have not yet been found outside the Lepidoptera, and are probably unique to the Lepidoptera. They show high sequence
and structural similarities to each other, indicating that the Lepidoptera-specific P450s may be of functional importance.
This analysis constitutes the first report of the clustering, spatial organization, and functional divergence of P450 in the
silkworm. 相似文献
4.
orf61 (bm61) of Bombyx mori Nucleopolyhedrovirus (BmNPV) is a highly conserved baculovirus gene, suggesting that it performs an important role in the
virus life cycle whose function is unknown. In this study, we describe the characterization of bm61. Quantitative polymerase chain reaction (qPCR) and western blot analysis demonstrated that bm61 was expressed as a late gene. Immunofluorescence analysis by confocal microscopy showed that BM61 protein was localized on
nuclear membrane and in intranuclear ring zone of infected cells. Structure localization of the BM61 in BV and ODV by western
analysis demonstrated that BM61 was the protein of both BV and ODV. In addition, our data indicated that BM61 was a late structure
protein localized in nucleus. 相似文献
5.
The mo (hereditary mosaic) mutation is one of the most famous and interesting mutations of the silkworm, Bombyx mori. Females homozygous for mo generate mosaic and gynandromorphic offspring due to non-elimination of polar bodies and subsequent double fertilization
events, irrespective of the genotype of the mated males. Although mo was first reported in 1927, the locus has not been mapped to linkage groups, as the mutation is unstable and appears to be
sensitive to genetic background. In this study, linkage analysis of mo was performed using PCR-based markers on single nucleotide polymorphism linkage maps. Bombyx mandarina was used as the mating partner for the B. mori
mo strain, as it is easier to identify polymorphic markers between B. mori and B. mandarina than within B. mori strains. Surprisingly, we identified two homozygous linkage groups (LGs) in all of the 12 B1 (first backcross generation) moths that had deposited mosaic eggs. It was revealed that +
mo
is located on the M chromosome of B. mandarina, which corresponds to two linkage groups of B. mori, LG 14 and 27. Based on further linkage analysis using B. mori as a mating partner, mo was mapped to LG 14. Additionally, we found that mo activity could be modified by a gene(s) on LG 17. 相似文献
6.
7.
Wolbachia are maternally inherited endosymbiotic bacteria of invertebrates that can manipulate the reproductive systems of their arthropod
hosts in a variety of ways. To establish a useful model system for investigating the mechanism of Wolbachia-induced host feminization, we conducted the following series of experiments: (1) feminizing Wolbachia of the butterfly, Eurema hecabe, were transferred into cell cultures of the silkmoth, Bombyx mori, and (2) the transfected Wolbachia in cell cultures were inoculated into B. mori at four immature stages. Wolbachia were successfully transfected into the cell cultures and stably maintained for more than 1 year (>30 passages). However,
none of the inoculated insects produced mature oocytes that were Wolbachia-positive. This finding was consistent with the fact that Wolbachia was not detected in individuals in subsequent generations. In contrast, Wolbachia were detected at relatively high frequencies (60–80% of individuals) in the somatic tissues of inoculated insects. Real-time
quantitative polymerase chain reaction revealed that the Wolbachia densities in the cultured cells were approximately tenfold higher than those in the native host E. hecabe. Among B. mori individuals inoculated at various developmental stages, those inoculated at early stages exhibited higher Wolbachia densities at the adult stage. The Wolbachia densities in individuals inoculated at the second-instar stage were comparable to those in intact E. hecabe. These results suggest that infection and/or proliferation of Wolbachia in germline cells are actively hindered by regulation in B. mori but feasible in somatic cells and that the Wolbachia densities in somatic tissues are regulated by the living host insects. 相似文献
8.
To investigate whether Bombyx mori immunized with Bacillus subtilis spore displaying GP64 escape from the B. mori nucleopolyhedrovirus (BmNPV) attack, a recombinant integrative plasmid named pJS700-GP64 was constructed, which carries a recombinant cotC-Gp64 gene under the control of the cotC promoter. In this study, pJS700-GP64 was transformed into B. subtilis 168 (trp−) competent cells, an amylase (amyE) inactivated mutant was selected, and was confirmed to be a double cross-over integrant, cotC-Gp64 fragment of which was integrated into B. subtilis chromosome. Gp64 was expressed on the spore surface and recognized by Gp64-specific antibody. Results of B. mori when challenged with BmNPV indicated that B. mori vaccinated with the recombinant spores possessed resistance to the invasion of BmNPV at some degree. 相似文献
9.
10.
WonKyung Kang 《中国病毒学》2009,24(4):315-322
Viruses including baculoviruses are obligatory parasites, as their genomes do not encode all the proteins required for replication.
Therefore, viruses have evolved to exploit the behavior and the physiology of their hosts and often coevolved with their hosts
over millions of years. Recent comparative analyses of complete genome sequences of baculoviruses revealed the patterns of
gene acquisitions and losses that have occurred during baculovirus evolution. In addition, knowledge of virus genes has also
provided understanding of the mechanism of baculovirus infection including replication, species-specific virulence and host
range. The Bm8 gene of Bombyx mori nucleopolyhedrovirus (NPV) and its homologues are found only in group I NPV genomes. The Autographa californica NPV Ac16 gene is a homologue of Bm8 and, encodes a viral structural protein. It has been shown that Bm8/Ac16 interacts with
baculoviral and cellular proteins. Bm8/Ac16 interacts with baculoviral IE1 that is facilitated by coiled coil domains, and
the interaction with IE1 is important for Bm8 function. Ac16 also forms a complex with viral FP25 and cellular actin and associates
with membranes via palmitoylation. These data suggested that this gene family encodes a multifunctional protein that accomplishes
specific needs of group I NPVs.
相似文献
11.
12.
Yi-Peng Xu Zheng-Pei Ye Chang-Ying Niu Yan-Yuan Bao Wen-Bing Wang Wei-De Shen Chuan-Xi Zhang 《Journal of microbiology (Seoul, Korea)》2010,48(1):102-110
The Bombyx mandarina nucleopolyhedrovirus (BomaNPV) S1 strain can infect the silkworm, Bombyx mori, but is significantly less virulent than B. mori nucleopolyhedrovirus (BmNPV) T3 strain. The complete nucleotide sequence of the S1 strain of BomaNPV was determined and compared
with the BmNPV T3 strain. The circular, double stranded DNA genome of the S1 strain was 126,770 nucleotides long (GenBank
accession no. FJ882854), with a G+C content of 40.23%. The genome contained 133 potential ORFs. Most of the putative proteins
were more than 96% identical to homologs in the BmNPV T3 strain, except for bro-a, lef-12, bro-c, and bro-d. Compared with the BmNPV T3 strain, however, this genome did not encode the bro-b and bro-e genes. In addition, hr1 lacked two repeat units, while hr2L, hr2R, hr3, hr4L, hr4R, and hr5 were similar to the corresponding
hrs in the T3 strain. The sequence strongly suggested that BomaNPV and BmNPV are variants with each other, and supported the
idea that baculovirus strain heterogeneity may often be caused by variation in the hrs and bro genes. 相似文献
13.
Xiao-long Hu Guang-li Cao Ren-yu Xue Xiao-jian Zheng Xing Zhang Hai-rong Duan Cheng-liang Gong 《Molecular biology reports》2010,37(6):2599-2608
The complete nucleotide sequence of the mitogenome of Bombyx mandarina strain Qingzhou was determined. The circular genome is 15,717 bp long and has the typical gene organization and order of
lepidopteran mitogenomes. All protein-coding sequences are initiated with a typical ATN codon, except the COI gene, which has a 4-bp TTAG putative initiator codon. Eleven of the 13 protein-coding gene have a complete termination codon
(all TAA), but the remaining two genes terminate with incomplete codons. All transfer RNAs (tRNAs) have a clover-leaf structure
typical of the mitochondrial tRNAs, and some of them have a mismatch in the four-stem-and-loop structure. The length of the
A + T rich region of B. mandarina strain Qingzhou is 495 bp, shorter than that of B. mandarina strain Tsukuba (747 bp) but similar to that of Bombyx mori. Phylogenetic analysis based on the whole mitochondrial genome sequences of the available sequenced species (B. mori strains C-108, Aojuku, Backokjam, and Xiafang, B. mandarina strains Tsukuba, Ankang, and Qingzhou, and Antheraea pernyi) shows the origin of the domesticated silkmoth B. mori to be the Chinese B. mandarina. Nuclear mitochondrial pseudogene sequences were detected in the nuclear genome of B. mori with the MEGA BLAST search program. A phylogenetic analysis of these nuclear mitochondrial pseudogene sequences suggests
that B. mori was domesticated independently in different areas and periods. 相似文献
14.
Shiping Liu Qingyou Xia Ping Zhao Tingcai Cheng Kaili Hong Zhonghuai Xiang 《BMC developmental biology》2007,7(1):88
Background
lin-4 and let-7, the two founding members of heterochronic microRNA genes, are firstly confirmed in Caenorhabditis elegans to control the proper timing of developmental programs in a heterochronic pathway. let-7 has been thought to trigger the onset of adulthood across animal phyla. Ecdysone and Broad-Complex are required for the temporal expression of let-7 in Drosophila melanogaster. For a better understanding of the conservation and functions of let-7, we seek to explore how it is expressed in the silkworm (Bombyx mori). 相似文献15.
Kawakami N Lee JM Mon H Kubo Y Banno Y Kawaguchi Y Maenaka K Park EY Koga K Kusakabe T 《Molecular biotechnology》2008,40(2):180-185
The recombinant protein expression by Bombyx mori nucleopolyhedrovirus (BmNPV) infecting silkworm larvae or pupae may endow us with a potent system for the production of large
eukaryotic proteins. However, the screening of silkworm strains ideally suited to this method has scarcely been conducted.
In the present study, we injected recombinant BmNPV containing a reporter gene, luciferase or DsRed, into hemocoel of fifth
instar larvae of selected 12 silkworm strains. Among them, the strain d17 is found to be the highest in reporter expression
from the intrinsic polyhedrin promoter of Autographa californica NPV or the silkworm actin A3 promoter. These results suggest that the d17 strain is highly permissive for BmNPV replication
and is the most likely candidate of a “factory” for large-scale expression using the BmNPV bacmid system. 相似文献
16.
Accurate quantification by real-time RT-PCR relies on normalisation of the measured gene expression data. Normalisation with
multiple reference genes is becoming the standard, but the best reference genes for gene expression studies within one organism
may depend on the applied treatments or the organs and tissues studied. Ideally, reference genes should be evaluated in all
experimental systems. A number of candidate reference genes for Arabidopsis have been proposed, which can be used as a starting point to evaluate their expression stability in individual experimental
systems by available computer algorithms like geNorm and NormFinder. Using this approach, we identified the best three reference
genes from a set of ten candidates, which included three traditional “housekeeping” genes, for normalisation of gene expression
when roots and leaves of Arabidopsis thaliana are exposed to cadmium (Cd) and copper (Cu). The expression stabilities of AT5G15710 (F-box protein), AT2G28390 (SAND family protein) and AT5G08290 (mitosis protein YLS8) were the highest when considering the effect to the roots and shoots of Cd and Cu treatments. Even
though the effect of Cd and excess Cu on the plants is very different, the same best reference genes were identified when
considering Cd or Cu treatments separately. This suggests that these three genes may also be suitable when studying the gene
expression after exposure of Arabidopsis thaliana to increased concentrations of other metals.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. 相似文献
17.
To construct the Bac-to-Bac expression system of Bombyx mori nucleopolyhedrovirus (BmNPV), a transfer vector was constructed which contained an Escherichia coli (E. coli) mini-F replicon and a lacZ: attTN7: lacZ cassette within the upstream and downstream regions of the BmNPV polyhedrin gene.
B. mori larvae were cotransfected with wild-type BmNPV genomic DNA and the transfer vector through subcutaneous injection to generate
recombinant viruses by homologous recombination in vivo. The genomic DNA of budded viruses extracted from the hemolymph of the transfected larvae was used to transform E. coli DH10B. Recombinant bacmids were screened by kanamycin resistance, PCR and restriction enzyme (REN) digestion. One of the
bacmid colonies, BmBacJS13, which had similar REN profiles to that of wild-type BmNPV, was selected for further research.
To investigate the infectivity of BmBacJS13, the polyhedrin gene was introduced into the bacmid and the resultant recombinant (BmBacJS13-ph) was transfected to BmN cells. The budded viruses were collected from the supernatant of the transfected cells and used for
infecting BmN cells. Growth curve analysis indicated that BmBacJS13-ph had a similar growth curve to that of wild-type BmNPV. Bio-assays indicated that BmBacJS13-ph was also infectious to B. mori larvae.
Foundation items: 973 (2003CB114202); Programme Strategic Scientific Alliances between China and the Netherlands (2004CB720404);
National Natural Fundation of China project (30630002) 相似文献
18.
Uchino K Imamura M Shimizu K Kanda T Tamura T 《Molecular genetics and genomics : MGG》2007,277(3):213-220
We investigated the use of Minos as a vector for transgenesis in the silkworm, Bombyx mori. We first constructed a vector plasmid with the green fluorescent protein (GFP) gene fused with the silkworm cytoplasmic
actin gene (A3) promoter, and a helper plasmid with the Minos transposase gene controlled by the same A3 promoter. Injection of the vector and helper plasmid DNA into silkworm eggs produced
transgenic animals in the following generation. The efficiency of transgenic silkworm production using this method was much
lower than that obtained using piggyBac-mediated germ line transformation. However, >40-fold increase in the efficiency of producing transgenic silkworms was obtained
using an in vitro synthesized source of Minos transposase mRNA. We conclude that the Minos transposon is a useful vector for construction of transgenic silkworms, particularly when in vitro synthesized mRNA is used.
This is the first report showing that Minos can be used as a vector for germ-line transformation in lepidopteran insects. 相似文献
19.
20.
Yasukochi Y Ashakumary LA Wu C Yoshido A Nohata J Mita K Sahara K 《Development genes and evolution》2004,214(12):606-614
A bacterial artificial chromosome (BAC) contig was constructed by chromosome walking, starting from the Hox genes of the silkworm, Bombyx mori. Bombyx orthologues of the labial (lab) and zerknült (zen) genes were newly identified. The size of the BAC contig containing the Hox gene cluster—except the lab and Hox 2 genes—was estimated to be more than 2 Mb. The Bombyx Hox cluster was mapped to linkage group (LG) 6. The lab gene was mapped on the same LG, but far apart from the cluster. Fluorescence in situ hybridization analysis confirmed that the major Hox gene cluster and lab were at different locations on the same chromosome in B. mori.Edited by M. Akam 相似文献