Abstract
Silk fibers have many inherent properties that are suitable for their use in biomaterials. In this study, the silk fibroin was genetically modified by including a Ca-binding sequence, [(AGSGAG)6ASEYDYDDDSDDDDEWD]2 from shell nacreous matrix protein. It can be produced as fibers by transgenic silkworm. The Ca-binding activity and mineralization of the transgenic silk fibroin were examined in vitro. The results showed that this transgenic silk fibroin had relatively higher Ca-binding activity than unmodified silk fibroin. The increased Ca-binding activity could promote the usage of silk fibroin as a biomaterial in the pharmaceutical industry. This study shows the possibility of using silk fibroin as a mineralization accelerating medical material by generating genetically modified transgenic silkworm.
Highlights
The silkworm Bombyx mori has been used as bioreactor to produce foreign proteins for decades due to its advanced physiological characteristics, such as short life cycle, convenient breeding process, large-scale protein production, and small individuals, which can be maintained in high densities, especially after the germline transformation method for the silkworm was developed using the transposon piggyBac[1,2,3,4]
The pBCABP-enhanced green fluorescent protein (EGFP) vector was injected into 1200 eggs of the Lan10 strain, and transgenic silkworms were screened for the expression of GFP in the whole body of G1 silkworms at the 1st instar under a fluorescence microscope equipped with a filter set for EGFP (Fig 2)
The results showed that there were no hybridizing bands identified in either type of sample, posterior silk glands and silk fibroin, at the target band molecular weight of 12.1 kDa
Summary
The silkworm Bombyx mori has been used as bioreactor to produce foreign proteins for decades due to its advanced physiological characteristics, such as short life cycle, convenient breeding process, large-scale protein production, and small individuals, which can be maintained in high densities, especially after the germline transformation method for the silkworm was developed using the transposon piggyBac[1,2,3,4]. Silk produced by silkworm is a natural protein fiber that contains two main components: fibroin and sericin. The clinical need of materials for bone regeneration is expected to increase, and some essential characteristics of these materials, including biocompatibility, porosity, and appropriate mechanical properties, are important for their application. The silk fibroin from silkworm, with superior mechanical properties such as the ability to be tailored, slow degradation, adequate time permitted for remodeling, and, most importantly, biocompatibility, is an ideal
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