scholarly journals A Targeted In-Fusion Expression System for Recombinant Protein Production in Bombyx mori

2022 ◽  
Vol 12 ◽  
Author(s):  
Zhiqian Li ◽  
Lang You ◽  
Qichao Zhang ◽  
Ye Yu ◽  
Anjiang Tan
Keyword(s):  
Bombyx Mori ◽  
Recombinant Proteins ◽  
Expression System ◽  
Silk Gland ◽  
Fusion Expression ◽  
Transcription Activator ◽  
Genomic Locus ◽  
Silk Proteins ◽  
Silk Glands ◽  
New Strategy

The domesticated silkworm, Bombyx mori, is an economically important insect that synthesizes large amounts of silk proteins in its silk gland to make cocoons. In recent years, germline transformation strategies advanced the bioengineering of the silk gland as an ideal bioreactor for mass production of recombinant proteins. However, the yield of exogenous proteins varied largely due to the random insertion and gene drift caused by canonical transposon-based transformation, calling for site-specific and stable expression systems. In the current study, we established a targeted in-fusion expression system by using the transcription activator-like effector nuclease (TALEN)-mediated targeted insertion to target genomic locus of sericin, one of the major silk proteins. We successfully generated chimeric Sericin1-EGFP (Ser-2A-EGFP) transformant, producing up to 3.1% (w/w) of EGFP protein in the cocoon shell. With this strategy, we further expressed the medically important human epidermal growth factor (hEGF) and the protein yield in both middle silk glands, and cocoon shells reached to more than 15-fold higher than the canonical piggyBac-based transgenesis. This natural Sericin1 expression system provides a new strategy for producing recombinant proteins by using the silkworm silk gland as the bioreactor.

scholarly journals Photoperiodic modulation of circadian rhythms in the silk gland protein profiles of Bombyx mori and its influence on the silk productivity and quality

2010 ◽  
Vol 2 (1) ◽  
pp. 48-56 ◽  
Author(s):  
B. Sailaja ◽  
S. Sivaprasad
Keyword(s):  
Circadian Rhythms ◽  
Bombyx Mori ◽  
Continuous Light ◽  
Silk Gland ◽  
Protein Profiles ◽  
Dark Cycle ◽  
Response Curves ◽  
Silk Proteins ◽  
Free Running ◽  
Clock Shift

Circadian rhythms in the silk gland protein profiles of Bombyx mori were analyzed under 12 h light and 12 h dark cycle (LD), continuous light (LL) and continuous dark (DD) conditions. The phase response curves of protein rhythms indicate the prevalence of a series of silk cycles, each comprising three phases; transcription, translation and consolidation of silk proteins. In the 24h- protein rhythm, the silk cycle repeats every 3h, 42 m under LD, 2h, 36m under LL and 3h under DD. The light and dark conditions advanced the rhythm of each silk cycle by 48m and 24m respectively. As a result the silk gland completes 7 rounds of protein synthesis under LD, 9 rounds under LL and 8 rounds under DD during the 24h-free running time of the rhythm. The light-induced clock-shift in the protein rhythm caused significant gains in economic parameters of sericulture with positive signals for enhancing silk productivity and quality.

TREHALASE ACTIVITY IN THE SILK GLANDS OF THE SILKWORM, BOMBYX MORI (LEPIDOPTERA: BOMBYCIDAE)

1975 ◽  
Vol 107 (12) ◽  
pp. 1311-1314 ◽  
Author(s):  
Shuya Shimada
Keyword(s):  
Bombyx Mori ◽  
Divalent Cations ◽  
Silk Gland ◽  
Trehalase Activity ◽  
Gland Cells ◽  
Silk Glands ◽  
Tunica Propria ◽  
Silkworm Bombyx Mori ◽  
Cu And Zn ◽  
Tunica Intima

Abstract(1) Crude extracts prepared from the silk glands of the silkworm, Bombyx mori L. contain trehalase activity. (2) Trehalase in the silk glands has a pH of 5.5 and a Km of 0.71 mM. The activity of the enzyme is inhibited by divalent cations such as Mn, Cu, and Zn. (3) By histochemical methods, it is shown that trehalase is localized in the periphery of the silk gland cells, especially in the tunica propria and tunica intima. (4) Trehalase activity is low in fifth instar and increases greatly in spinning stages, after which the activity decreases.

An optimized sericin-1 expression system for mass-producing recombinant proteins in the middle silk glands of transgenic silkworms

2013 ◽  
Vol 22 (5) ◽  
pp. 925-938 ◽  
Author(s):  
Feng Wang ◽  
Hanfu Xu ◽  
Lin Yuan ◽  
Sanyuan Ma ◽  
Yuancheng Wang ◽  
...  
Keyword(s):  
Recombinant Proteins ◽  
Expression System ◽  
Silk Glands ◽  
Transgenic Silkworms

scholarly journals Reference transcriptome data in silkworm Bombyx mori

2019 ◽  
Author(s):  
Kakeru Yokoi ◽  
Takuya Tsubota ◽  
Jianqiang Sun ◽  
Akiya Jouraku ◽  
Hideki Sezutsu ◽  
...  
Keyword(s):  
Bombyx Mori ◽  
Reference Genome ◽  
Enrichment Analysis ◽  
Silk Gland ◽  
Transcriptome Data ◽  
Data Set ◽  
Reference Transcriptome ◽  
Genome Data ◽  
Silk Glands ◽  
Sequence Region

AbstractThe silkworm Bombyx mori has long been used in the silk industry and utilized in studies of physiology, genetics, molecular biology, and pathology. We recently reported high quality reference genome data for B. mori. In the present study, we constructed a reference transcriptome data set using the reference genome data and RNA-seq data of 10 tissues from P50T strain larvae. Reference transcriptome data contained 51,926 transcripts, with 39,619 having one or more coding sequence region. The abundance of each transcript in the 10 tissues as well as 5 tissues from other strain larvae was estimated, and hierarchical clustering was performed. The results obtained showed that data on abundance were highly reproducible and there here is a little difference of transcriptome abundance between the two strain larvae. New isoforms of silk-related genes were searched for in the reference transcriptomes, and the longest isoform of sericin-1 possessing a long exon was identified. We also extracted transcripts that were strongly expressed in one or more parts of the silk glands. An enrichment analysis performed using the functional annotation data of the transcripts provided novel insights into the functions of the silk gland parts. Therefore, the reference transcriptome data set obtained has extended B. mori genomic and transcriptomic insights and may contribute to advances in entomologic and vertebrate research, including that on humans.

Expression of autophagy-related genes in the anterior silk gland of the silkworm (Bombyx mori) during metamorphosis

2011 ◽  
Vol 89 (11) ◽  
pp. 1019-1026 ◽  
Author(s):  
Qingrong Li ◽  
Xiaojuan Deng ◽  
Zhijun Huang ◽  
Sichun Zheng ◽  
Gianluca Tettamanti ◽  
...  
Keyword(s):  
Bombyx Mori ◽  
Morphological Characteristics ◽  
Time Frame ◽  
Silk Gland ◽  
Marker Genes ◽  
Reverse Transcription Pcr ◽  
Evolutionarily Conserved ◽  
Silk Glands ◽  
Autophagic Process ◽  
Silkworm Bombyx Mori

Programmed cell death (PCD) is a genetically regulated process of cell elimination and is evolutionarily conserved in eukaryotes. Degeneration of larval tissues during metamorphosis in insects is a result of PCD triggered by ecdysteroids and autophagic process has been shown to be involved in the degeneration of silk gland of the silkworm ( Bombyx mori L., 1758). However, experimental evidence for the expression of autophagy marker genes remains insufficient. In this study, expression of the autophagy-related genes BmAtg5, BmAtg6, BmAtg8, and BmAtg12 was determined in the anterior silk glands of larvae and pupae during larval to pupal transformation by using reverse-transcription PCR, quantitative real-time PCR, and Western blot analyses. The results indicated that BmAtg5, BmAtg6, BmAtg8, and BmAtg12 had the highest expression levels on the 3rd day of the wandering stage or at the prepupal stage, and the time frame of the expression of these genes was coincident with the morphological characteristics of autophagy in the silk gland during the larval to pupal transformation. This study demonstrated that the autophagy-related genes are involved in the degeneration of the anterior silk gland of B. mori.

A novel, cheap and effective fusion expression system for the production of recombinant proteins

2007 ◽  
Vol 77 (2) ◽  
pp. 483-488 ◽  
Author(s):  
Fei-Xiang Ding ◽  
Hong-Li Yan ◽  
Qian Mei ◽  
Geng Xue ◽  
Yu-Zhao Wang ◽  
...  
Keyword(s):  
Recombinant Proteins ◽  
Expression System ◽  
Fusion Expression

Role of Bm30kc6 gene in cell apoptosis and the silk gland degradation signaling pathway in Bombyx mori L.

2020 ◽  
Vol 105 (3) ◽  
Author(s):  
Ying Xiao ◽  
Lei‐lei Li ◽  
Asma Bibi ◽  
Ning Zhang ◽  
Ting Chen ◽  
...  
Keyword(s):  
Bombyx Mori ◽  
Signaling Pathway ◽  
Cell Apoptosis ◽  
Silk Gland ◽  
Bombyx Mori L

scholarly journals Applicability of Honey on Silkworms (Bombyx mori) and Quality Improvement of Its Biomaterials

2021 ◽  
Vol 11 (10) ◽  
pp. 4613
Author(s):  
Gabriela-Maria Baci ◽  
Alexandra-Antonia Cucu ◽  
Adela Ramona Moise ◽  
Daniel Severus Dezmirean
Keyword(s):  
Quality Improvement ◽  
Bombyx Mori ◽  
Human Health ◽  
Recombinant Proteins ◽  
Therapeutic Agent ◽  
Life Sciences ◽  
Model Organism ◽  
Medical Field ◽  
Wide Range ◽  
Ancient Times

Since ancient times, honey has been considered one of the most illustrious and esteemed natural products. Honey plays two key roles; specifically, it is an appreciated nutritional product, and also exhibits a wide range of beneficial properties for human health as a therapeutic agent. Furthermore, it has been shown that honey has valuable effects on the biological and physiological features of mulberry silkworms (Bombyx mori). Bombyx mori exhibits importance not only for the economy, but it also serves as an important biotechnological bioreactor for the production of recombinant proteins that have a great impact in the medical field and beyond. It also represents an important model organism for life sciences. In view of the fact that silk fibroin serves as a natural biopolymer that displays high biocompatibility with human organisms and due to honey’s various and remarkable properties for human health, the two elements are currently used together in order to develop ideal biomaterials for a wide range of purposes. In this review, by discussing the applicability of honey on Bombyx mori and beyond, the importance of honey for life sciences and related fields is spotlighted.

scholarly journals Bombyx mori Pupae Efficiently Produce Recombinant AAV2/HBoV1 Vectors with a Bombyx mori Nuclear Polyhedrosis Virus Expression System

Viruses ◽  
2021 ◽  
Vol 13 (4) ◽  
pp. 704
Author(s):  
Qian Yu ◽  
Pengfei Chang ◽  
Xiaoxuan Liu ◽  
Peng Lü ◽  
Qi Tang ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Bombyx Mori ◽  
Nuclear Polyhedrosis Virus ◽  
Expression System ◽  
Efficient Production ◽  
Human Bocavirus ◽  
Adeno Associated Virus ◽  
Silkworm Pupae ◽  
Polyhedrosis Virus ◽  
Nuclear Polyhedrosis

Recombinant adeno-associated virus (AAV) vectors have broad application prospects in the field of gene therapy. The establishment of low-cost and large-scale manufacturing is now the general agenda for industry. The baculovirus-insect cell/larva expression system has great potential for these applications due to its scalability and predictable biosafety. To establish a more efficient production system, Bombyx mori pupae were used as a new platform and infected with recombinant Bombyx mori nuclear polyhedrosis virus (BmNPV). The production of a chimeric recombinant adeno-associated virus (rAAV) serotype 2/human bocavirus type-1 (HBoV1) vector was used to evaluate the efficiency of this new baculovirus expression vector (BEV)–insect expression system. For this purpose, we constructed two recombinant BmNPVs, which were named rBmNPV/AAV2Rep-HBoV1Cap and rBmNPV/AAV2ITR-eGFP. The yields of rAAV2/HBoV1 derived from the rBmNPV/AAV2Rep-HBoV1Cap and rBmNPV/AAV2ITR-eGFP co-infected BmN cells exceeded 2 × 104 vector genomes (VG) per cell. The rBmNPV/AAV2Rep-HBoV1Cap and rBmNPV/AAV2ITR-eGFP can express stably for at least five passages. Significantly, rAAV2/HBoV1 could be efficiently generated from BmNPV-infected silkworm larvae and pupae at average yields of 2.52 × 1012 VG/larva and 4.6 × 1012 VG/pupa, respectively. However, the vectors produced from the larvae and pupae had a high percentage of empty particles, which suggests that further optimization is required for this platform in the future. Our work shows that silkworm pupae, as an efficient bioreactor, have great potential for application in the production of gene therapy vectors.

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