PDF(Pubmed)
Abstract：
UNASSIGNED: Lucilia sericata (Diptera: Calliphoridae) is used in larval therapy for wound healing. Netrin-A is an enzyme secreted from the salivary glands of these larvae, and has a central role in neural regeneration and angiogenesis. This study aimed to produce the recombinant Netrin-A protein from Lucilia sericata larvae by the baculovirus expression vector system in the Sf9 insect cell line.
UNASSIGNED: The coding sequence of Netrin-A was cloned, amplified in the pTG19 vector, and then cloned in the pFastBac HTA vector. It was then transformed into DH10Bac, and the recombinant Bacmid was subsequently transfected into Sf9 cells. The recombinant Netrin-A was purified by Ni-NTA agarose. The evaluation was done using SDS-PAGE and western blot, respectively. Finally, its concentration was calculated with the Bradford assay.
UNASSIGNED: The molecular weight of this protein was 52 kDa with 404 amino acids. The signal peptide was located between amino acids 24 and 25. The concentration of Netrin-A was calculated to be 48.8 μg/ml. It reaffirmed the characterized gene codes of Lucilia sericata Netrin-A in a previous study.
UNASSIGNED: The generation of recombinant Netrin-A could be used in larval therapy, and as a biomarker in certain diseases. The netrin-A of Lucilia sericata was unprecedentedly cloned and expressed in a eukaryotic cell line. Given that this larva is FDA-approved, and non-pathogenic, it conduces to research on the development of maggot therapy in future.
UNASSIGNED: The coding sequence of Netrin-A was cloned, amplified in the pTG19 vector, and then cloned in the pFastBac HTA vector. It was then transformed into DH10Bac, and the recombinant Bacmid was subsequently transfected into Sf9 cells. The recombinant Netrin-A was purified by Ni-NTA agarose. The evaluation was done using SDS-PAGE and western blot, respectively. Finally, its concentration was calculated with the Bradford assay.
UNASSIGNED: The molecular weight of this protein was 52 kDa with 404 amino acids. The signal peptide was located between amino acids 24 and 25. The concentration of Netrin-A was calculated to be 48.8 μg/ml. It reaffirmed the characterized gene codes of Lucilia sericata Netrin-A in a previous study.
UNASSIGNED: The generation of recombinant Netrin-A could be used in larval therapy, and as a biomarker in certain diseases. The netrin-A of Lucilia sericata was unprecedentedly cloned and expressed in a eukaryotic cell line. Given that this larva is FDA-approved, and non-pathogenic, it conduces to research on the development of maggot therapy in future.
摘要：
■丝龙（双翅目：Calliphoridae）用于幼虫治疗伤口愈合。Netrin-A是从这些幼虫的唾液腺分泌的一种酶，在神经再生和血管生成中起着核心作用。本研究旨在通过杆状病毒表达载体系统，在Sf9昆虫细胞系中从丝黄幼虫中生产重组Netrin-A蛋白。
■克隆了Netrin-A的编码序列，在pTG19载体中扩增，然后克隆到pFastBacHTA载体中。然后转化为DH10Bac，然后将重组Bacmid转染到Sf9细胞中。通过Ni-NTA琼脂糖纯化重组Netrin-A。使用SDS-PAGE和蛋白质印迹进行评估，分别。最后,用Bradford测定法计算其浓度。
■该蛋白的分子量为52kDa,具有404个氨基酸。信号肽位于氨基酸24和25之间。Netrin-A的浓度计算为48.8μg/ml。在先前的研究中，它重申了LuciliasericataNetrin-A的特征基因密码。
■重组Netrin-A的产生可用于幼虫治疗，并作为某些疾病的生物标志物。sericata的netrin-A被前所未有地克隆并在真核细胞系中表达。鉴于这种幼虫是FDA批准的，和非致病性，它有助于研究未来的虫疗法的发展。
■克隆了Netrin-A的编码序列，在pTG19载体中扩增，然后克隆到pFastBacHTA载体中。然后转化为DH10Bac，然后将重组Bacmid转染到Sf9细胞中。通过Ni-NTA琼脂糖纯化重组Netrin-A。使用SDS-PAGE和蛋白质印迹进行评估，分别。最后,用Bradford测定法计算其浓度。
■该蛋白的分子量为52kDa,具有404个氨基酸。信号肽位于氨基酸24和25之间。Netrin-A的浓度计算为48.8μg/ml。在先前的研究中，它重申了LuciliasericataNetrin-A的特征基因密码。
■重组Netrin-A的产生可用于幼虫治疗，并作为某些疾病的生物标志物。sericata的netrin-A被前所未有地克隆并在真核细胞系中表达。鉴于这种幼虫是FDA批准的，和非致病性，它有助于研究未来的虫疗法的发展。
