A review on complete silk gene sequencing and de novo assembly of artificial silk.

Silk, especially spider and insect silk, is a highly versatile biomaterial with potential applications in biomedicine, materials science, and biomimetic engineering. The primary structure of silk proteins is the basis for the mechanical properties of silk fibers. Biotechnologies such as single-molecule sequencing have facilitated an increasing number of reports on new silk genes and assembled silk proteins. Therefore, this review aims to provide a comprehensive overview of the recent advances in representative spider and insect silk proteins, focusing on identification methods, sequence characteristics, and de novo design and assembly. The review discusses three identification methods for silk genes: polymerase chain reaction (PCR)-based sequencing, PCR-free cloning and sequencing, and whole-genome sequencing. Moreover, it reveals the main spider and insect silk proteins and their sequences. Subsequent de novo assembly of artificial silk is covered and future research directions in the field of silk proteins, including new silk genes, customizable artificial silk, and the expansion of silk production and applications are discussed. This review provides a basis for the genetic aspects of silk production and the potential applications of artificial silk in material science and biomedical engineering.

Non-classical digestive lipase BmTGL selected by gene amplification reduces the effects of mulberry inhibitor during silkworm domestication.

Efficient utilization of dietary lipids is crucial for Bombyx mori, also known as domesticated silkworms. However, the effects of domestication on the genes encoding lipases remain unknown. In this study, we investigated the expression difference of one triacylglycerol lipase (BmTGL) between B.mori and wild (ancestor) silkworm strains (Bombyx mandarina). An immunofluorescence localization analysis showed that BmTGL was present in all parts of the gut and was released into the intestinal lumen. BmTGL expression was significantly enhanced in different domesticated silkworm strains compared to that in the B. mandarina strains. The BmTGL copy numbers in the genomes of the domesticated silkworm strains were 2-to-3 folds that of the B. mandarina strains and accounted for the enhanced expression of BmTGL in the domesticated silkworm strains. The Ser144Asn substitution in the Ser-Asp-His catalytic triads of BmTGL resulted in relatively lower lipase activity and reduced sensitivity to the lipase inhibitor morachalcone A. Moreover, BmTGL overexpression significantly increased the weights of the B. mori silkworms compared to those of the non-transgenic controls. Thus, the selection of BmTGL by gene amplification may be a trade-off between maintaining high enzymatic activity and reducing the effects of mulberry inhibitors during silkworm domestication.

Microstructure Evolution Mechanism of Ultra-Thin Dual Phase Magnesium-Lithium Alloy during Asymmetric Warm Rolling.

Magnesium-lithium alloy is the lightest metal alloy material so far, and the ultra-thin plate is also one of the main trends in the future development of Mg-Li alloy. In order to explore how to prepare LZ91 ultra-thin Mg-Li alloy, this topic adopts the combination of the finite element method (FEM) and visco-plastic self-consistent (VPSC) calculation, electron back-scattered diffraction (EBSD) and tensile experiment, and uses the asymmetric warm rolling process to realize the processing of ultra-thin LZ91 Mg-Li alloy plate with a thickness of 0.25 mm. The experimental results show that the maximum basal texture strengths of 1 mm initial plate and 0.25 mm ultra-thin rolled plate are 36.02 mud and 29.19 mud, respectively. The asymmetric warm rolling process not only reduces the basal texture strength but also significantly refines the grains. The tensile strength and yield strength of 0.25 mm ultra-thin rolled plate along the rolling direction reached 206.8 MPa and 138.4 MPa, respectively. This has a positive effect on the mechanical properties of subsequent materials. VPSC results show that the base slip is the main factor in Mg-Li alloy asymmetric warm rolling, and a large number of tensile twinning are initiated due to the coordinated deformation of the body-centered cubic (BCC) phase, which is beneficial to improve the plastic deformation capacity of Mg-Li alloy.

Biochemical characterization and overexpression of an α-amylase (BmAmy) in silkworm, Bombyx mori.

Silkworm (Bombyx mori) is the only fully domesticated insect. As an economically important insect, nutrition utilization is important for its productivity. Hence, the present study investigated the expression pattern of BmAmy, an α-amylase, in B. mori. BmAmy protein purification and biochemical characterization were performed, and effects of BmAmy overexpression were assessed. Real-time quantitative reverse transcription polymerase chain reaction indicated that BmAmy transcription was positively correlated with the silkworm's food intate. Moreover, enzymatic activity assay results showed that BmAmy had significant α-amylase activity of about 1 mg/min/mg protein. Furthermore, treatment with mulberry amylase inhibitors MnAI1 and MnAI2 resulted to 89.92% and 93.67% inhibition in BmAmy activity, respectively, and the interaction between BmAmy and MnAI was also confirmed by protein docking analysis. A silkworm line that specifically overexpressed BmAmy in the midgut was generated through piggyBac-based transgenic technology, and compared to those of non-transgenic silkworms, the whole cocoon and cocoon shell weights of these transgenic silkworms increased by 10.13% and 18.32%, respectively, in the female group, and by 5.83% and 6.00%, respectively, in the male group. These results suggested that BmAmy may be a suitable target for breeding better silkworm varieties in the future.