A silkworm that can produce a chimeric silk thread in fibrous form with physical properties which approximate those of the bagworm silk thread is provided. A genome-modified silkworm is also provided in which a gene encoding a modified bagworm fibroin protein is inserted, wherein the gene is obtained by fusing a gene fragment encoding a bagworm-derived fibroin protein and a gene fragment encoding a silkworm-derived fibroin protein.

Disclosed is a silk gland recombinant expression vector of silkworm expressing human epidermal growth factor, a preparation method and use thereof. The recombinant expression vector is formed by fusing truncated silk fibroin light chain N-terminal domain, C-terminal domain, optimized human epidermal growth factor and silkworm nuclear polyhedrosis virus enhancer, and removes the redundant part of the extra domain of endogenous silk protein contained in the fusion protein to the maximum extent.

The application describes methods for synthetic synthesis and cell-free synthesis of DNA vectors, particularly closed-ended DNA vectors (e.g., ceDNA vectors) having linear and continuous structure for delivery and expression of a transgene. The present invention relates to an in vitro process for production of closed-ended DNA vectors, corresponding DNA vector products produced by the methods and uses thereof, and oligonucleotides and kits useful in the process of the invention. DNA vectors produced using the methods described herein are free from unwanted side effects due to contaminants introduced during production in cell lines, for example, bacterial or insect cell lines. Further provided herein are methods and cell lines for reliable gene expression in vitro, ex vivo and in vivo using the ceDNA vectors synthesized using the methods herein.

The disclosure discloses an expression cassette for expressing a gene including overlapping open reading frames in an insect cell and an application thereof. The expression cassette includes from 5 to 3 and operably linked: a promoter capable of driving transcription in the insect cell; an artificially constructed sequence; the overlapping open reading frames missing only a first translation start codon; wherein the artificially constructed sequence includes a native or engineered intron with splicing activity in the insect cell, the intron includes ATG or the intron is located between any two adjacent nucleotides in ATG. A recombinant adeno-associated virus vector including the expression cassette of the disclosure regulates relative expressions of VP1, VP2, and VP3 proteins, and relative expressions of Rep78 and Rep52 proteins by using a designed intron sequence and through an intron splicing function for large-scale production of rAAV.

Disclosed is a silk gland recombinant expression vector of silkworm expressing human epidermal growth factor, a preparation method and use thereof. The recombinant expression vector is formed by fusing truncated silk fibroin light chain N-terminal domain, C-terminal domain, optimized human epidermal growth factor and silkworm nuclear polyhedrosis virus enhancer, and removes the redundant part of the extra domain of endogenous silk protein contained in the fusion protein to the maximum extent.

The invention relates to identification and characterization of recombinant DNA and polypeptides for specific Bt toxin receptors. In particular, the Bt toxin receptors of the invention include those derived from the Lepidopteran super family including the species Trichoplusiani ni, Pseudoplusia includens, Helicoverpa zea, and Spodoptera frugiperda. The receptors of the invention further include those derived from the Coleopteran super family and particularly from the species Diabrotica virgifera virgifera. The recombinant DNA and polypeptides so provided are useful in the identification and design of novel Bt toxin receptor ligands including novel or improved insecticidal toxins for use in a variety of agricultural applications. Materials and methods for identifying novel toxins are also disclosed herein. The invention also provides methods for selecting toxins to combine to control insect populations by manipulating Bt toxin receptor.