title: ÐÑÑÐ»ÐµÐ´Ð¾Ð²Ð°Ð½Ð¸Ðµ Ð²Ð»Ð¸ÑÐ½Ð¸Ñ Ð¼Ð¾Ð´Ð¸ÑÐ¸ÐºÐ°ÑÐ¸Ð¸ Ð³ÐµÐ½Ð° L1 Ð¿Ð°Ð¿Ð¸Ð»Ð»Ð¾Ð¼Ð°Ð²Ð¸ÑÑÑÐ° HPV16 Ð½Ð° ÑÐ°ÑÑÐ²Ð¾ÑÐ¸Ð¼Ð¾ÑÑÑ Ð¿ÑÐ¾Ð´ÑÐºÑÐ° ÐµÐ³Ð¾ ÑÐºÑÐ¿ÑÐµÑÑÐ¸Ð¸
title: Ð²ÑÐ¿ÑÑÐºÐ½Ð°Ñ ÐºÐ²Ð°Ð»Ð¸ÑÐ¸ÐºÐ°ÑÐ¸Ð¾Ð½Ð½Ð°Ñ ÑÐ°Ð±Ð¾ÑÐ° Ð±Ð°ÐºÐ°Ð»Ð°Ð²ÑÐ°
titleType: Subtitle
title: Study of the influence of modifications of the L1 gene of the HPV16 papillomavirus on the solubility of its expression product
Creators: name: Ð¢ÐºÐ°ÑÐµÐ²Ð°, ÐÐ°ÑÐ¸Ð½Ð°
Publisher: name: Ð¡Ð°Ð½ÐºÑ-ÐÐµÑÐµÑÐ±ÑÑÐ³ÑÐºÐ¸Ð¹ Ð¿Ð¾Ð»Ð¸ÑÐµÑÐ½Ð¸ÑÐµÑÐºÐ¸Ð¹ ÑÐ½Ð¸Ð²ÐµÑÑÐ¸ÑÐµÑ ÐÐµÑÑÐ° ÐÐµÐ»Ð¸ÐºÐ¾Ð³Ð¾
Year: 2024
Subjects: Ð²Ð¸ÑÑÑ Ð¿Ð°Ð¿Ð¸Ð»Ð»Ð¾Ð¼Ñ ÑÐµÐ»Ð¾Ð²ÐµÐºÐ°
Ð¼Ð¾Ð´Ð¸ÑÐ¸ÐºÐ°ÑÐ¸Ð¸ Ð³ÐµÐ½Ð° L1 Ð¿Ð°Ð¿Ð¸Ð»Ð»Ð¾Ð¼Ð°Ð²Ð¸ÑÑÑÐ° HPV16
Ð±ÐµÐ»Ð¾Ðº L1
ÑÐ²ÐµÐ»Ð¸ÑÐµÐ½Ð¸Ðµ ÑÐ°ÑÑÐ²Ð¾ÑÐ¸Ð¼Ð¾ÑÑÐ¸ Ð±ÐµÐ»ÐºÐ°
ÑÐ»Ð¸ÑÑÐµ Ð±ÐµÐ»ÐºÐ¸
human papilloma virus
HPV16 papillomavirus L1 gene modifications
L1 protein
increased protein solubility
fused proteins

description: ÐÐ°Ð½Ð½Ð°Ñ ÑÐ°Ð±Ð¾ÑÐ° Ð¿Ð¾ÑÐ²ÑÑÐµÐ½Ð° Ð¿Ð¾Ð»ÑÑÐµÐ½Ð¸Ñ ÑÐ°ÑÑÐ²Ð¾ÑÐ¸Ð¼ÑÑ Ð±ÐµÐ»ÐºÐ¾Ð² L1 HPV16 Ð´Ð»Ñ Ð´Ð°Ð»ÑÐ½ÐµÐ¹ÑÐµÐ¹ ÑÐ°Ð·ÑÐ°Ð±Ð¾ÑÐºÐ¸ ÐÐ¤Ð ÑÐµÑÑ-ÑÐ¸ÑÑÐµÐ¼Ñ. ÐÐ°Ð´Ð°ÑÐ¸, ÐºÐ¾ÑÐ¾ÑÑÐµ ÑÐµÑÐ°Ð»Ð¸ÑÑ Ð² ÑÐ¾Ð´Ðµ Ð¸ÑÑÐ»ÐµÐ´Ð¾Ð²Ð°Ð½Ð¸Ñ: 1. ÐÑÐ¾Ð²ÐµÑÑÐ¸ Ð¾Ð±Ð·Ð¾Ñ Ð»Ð¸ÑÐµÑÐ°ÑÑÑÑ Ð½Ð° Ð¸ÑÑÐ»ÐµÐ´ÑÐµÐ¼ÑÑ ÑÐµÐ¼Ñ. 2. Ð¡Ð¾Ð·Ð´Ð°ÑÑ Ð³ÐµÐ½ÐµÑÐ¸ÑÐµÑÐºÐ¸Ðµ ÐºÐ¾Ð½ÑÑÑÑÐºÑÐ¸Ð¸ Ñ Ð¼Ð¾Ð´Ð¸ÑÐ¸ÐºÐ°ÑÐ¸ÑÐ¼Ð¸ Ð³ÐµÐ½Ð° L1. 3. ÐÑÐ¾Ð²ÐµÑÐ¸ÑÑ Ð²Ð»Ð¸ÑÐ½Ð¸Ðµ ÑÐºÐ¾ÑÐ¾ÑÐµÐ½Ð¸Ñ Ð³ÐµÐ½Ð° Ð±ÐµÐ»ÐºÐ° L1 Ð½Ð° ÐµÐ³Ð¾ ÑÐ°ÑÑÐ²Ð¾ÑÐ¸Ð¼Ð¾ÑÑÑ. 4. Ð¡Ð¾Ð·Ð´Ð°ÑÑ Ð³ÐµÐ½ÐµÑÐ¸ÑÐµÑÐºÐ¸Ðµ ÐºÐ¾Ð½ÑÑÑÑÐºÑÐ¸Ð¸ Ð¼Ð¾Ð´Ð¸ÑÐ¸ÑÐ¸ÑÐ¾Ð²Ð°Ð½Ð½Ð¾Ð³Ð¾ Ð±ÐµÐ»ÐºÐ° L1 Ñ ÑÑÐ³Ð°Ð¼Ð¸. 5. ÐÑÐ¾Ð²ÐµÑÐ¸ÑÑ Ð²Ð»Ð¸ÑÐ½Ð¸Ðµ ÑÑÐ³Ð¾Ð² Ð½Ð° ÑÐ°ÑÑÐ²Ð¾ÑÐ¸Ð¼Ð¾ÑÑÑ Ð¼Ð¾Ð´Ð¸ÑÐ¸ÑÐ¸ÑÐ¾Ð²Ð°Ð½Ð½Ð¾Ð³Ð¾ Ð±ÐµÐ»ÐºÐ° L1. ÐÐ±ÑÐµÐºÑ Ð¸ÑÑÐ»ÐµÐ´Ð¾Ð²Ð°Ð½Ð¸Ñ â Ð±ÐµÐ»Ð¾Ðº L1 Ð¿Ð°Ð¿Ð¸Ð»Ð»Ð¾Ð¼Ð°Ð²Ð¸ÑÑÑÐ° HPV16. ÐÑÐµÐ´Ð¼ÐµÑ Ð¸ÑÑÐ»ÐµÐ´Ð¾Ð²Ð°Ð½Ð¸Ñ â ÑÐ°ÑÑÐ²Ð¾ÑÐ¸Ð¼Ð¾ÑÑÑ Ð±ÐµÐ»ÐºÐ° L1. Ð Ð°Ð±Ð¾ÑÐ° Ð²ÑÐ¿Ð¾Ð»Ð½ÐµÐ½Ð° Ð½Ð° Ð±Ð°Ð·Ðµ Ð»Ð°Ð±Ð¾ÑÐ°ÑÐ¾ÑÐ¸Ð¸ Ð¼Ð¾Ð»ÐµÐºÑÐ»ÑÑÐ½Ð¾Ð¹ Ð³ÐµÐ½ÐµÑÐ¸ÐºÐ¸ Ð¿Ð°ÑÐ¾Ð³ÐµÐ½Ð½ÑÑ Ð¼Ð¸ÐºÑÐ¾Ð¾ÑÐ³Ð°Ð½Ð¸Ð·Ð¼Ð¾Ð² Ð¤ÐÐ£Ð ÐÐÐÐ­Ð Ð¸Ð¼ÐµÐ½Ð¸ ÐÐ°ÑÑÐµÑÐ°. ÐÑÐ»Ð¸ ÑÐ¾Ð±ÑÐ°Ð½Ñ 3 Ð³ÐµÐ½ÐµÑÐ¸ÑÐµÑÐºÐ¸Ðµ ÐºÐ¾Ð½ÑÑÑÑÐºÑÐ¸Ð¸ Ð¼ÐµÑÐ¾Ð´Ð¾Ð¼ ÑÐµÑÑÑÐ¸ÐºÑÐ¸Ð¸-Ð»Ð¸Ð³Ð¸ÑÐ¾Ð²Ð°Ð½Ð¸Ñ Ñ ÑÐ°Ð·Ð½ÑÐ¼Ð¸ Ð²Ð°ÑÐ¸Ð°Ð½ÑÐ°Ð¼Ð¸ Ð¼Ð¾Ð´Ð¸ÑÐ¸ÐºÐ°ÑÐ¸Ð¸ Ð³ÐµÐ½Ð° L1, Ð° ÑÐ°ÐºÐ¶Ðµ 9 Ð³ÐµÐ½ÐµÑÐ¸ÑÐµÑÐºÐ¸Ñ ÐºÐ¾Ð½ÑÑÑÑÐºÑÐ¸Ð¹ Ñ Ð´Ð¾Ð¿Ð¾Ð»Ð½Ð¸ÑÐµÐ»ÑÐ½ÑÐ¼Ð¸ ÑÐµÐ³Ð°Ð¼Ð¸: MBP, GST, TF Ð¸ Ð¼Ð¾Ð´Ð¸ÑÐ¸ÑÐ¸ÑÐ¾Ð²Ð°Ð½Ð½ÑÐ¼ Ð³ÐµÐ½Ð¾Ð¼ L1. ÐÐ»Ñ Ð°Ð½Ð°Ð»Ð¸Ð·Ð° ÑÑÑÐµÐºÑÐ¸Ð²Ð½Ð¾ÑÑÐ¸ Ð½Ð°ÑÐ°Ð±Ð¾ÑÐºÐ¸ Ð±ÐµÐ»ÐºÐ°, Ð° ÑÐ°ÐºÐ¶Ðµ Ð²ÑÑÐ²Ð»ÐµÐ½Ð¸Ñ ÑÐ°ÑÑÐ²Ð¾ÑÐ¸Ð¼Ð¾ÑÑÐ¸ Ð±ÐµÐ»ÐºÐ° Ð¸Ð½ÑÐµÑÐµÑÐ° Ð±ÑÐ»Ð¸ Ð¿ÑÐ¾Ð²ÐµÐ´ÐµÐ½Ñ ÑÐ»ÐµÐ´ÑÑÑÐ¸Ðµ ÑÑÐ°Ð¿Ñ: ÐºÑÐ»ÑÑÐ¸Ð²Ð¸ÑÐ¾Ð²Ð°Ð½Ð¸Ðµ, Ð¿Ð¾Ð´Ð³Ð¾ÑÐ¾Ð²ÐºÐ° ÑÑÐ°ÐºÑÐ¸Ð¹ Ð¸ Ð¿ÑÐ¾Ð± Ð´Ð»Ñ ÑÐ»ÐµÐºÑÑÐ¾ÑÐ¾ÑÐµÐ·Ð° Ð±ÐµÐ»ÐºÐ° Ð² Ð¿Ð¾Ð»Ð¸Ð°ÐºÑÐ¸Ð»Ð°Ð¼Ð¸Ð´Ð½Ð¾Ð¼ Ð³ÐµÐ»Ðµ Ð² Ð¿ÑÐ¸ÑÑÑÑÑÐ²Ð¸Ð¸ Ð´Ð¾Ð´ÐµÑÐ¸Ð»ÑÑÐ»ÑÑÐ°ÑÐ° Ð½Ð°ÑÑÐ¸Ñ Ð¿Ð¾ Ð¼ÐµÑÐ¾Ð´Ñ ÐÑÐ¼Ð¼Ð»Ð¸. Ð ÑÐ¾Ð´Ðµ Ð¸ÑÑÐ»ÐµÐ´Ð¾Ð²Ð°Ð½Ð¸Ñ Ð²ÑÑÐ²Ð»ÐµÐ½Ð¾, ÑÑÐ¾ ÑÐºÐ¾ÑÐ¾ÑÐµÐ½Ð¸Ðµ Ð³ÐµÐ½Ð° Ð½Ðµ ÑÐ¾Ð´ÐµÐ¹ÑÑÐ²ÑÐµÑ ÑÐ²ÐµÐ»Ð¸ÑÐµÐ½Ð¸Ñ ÑÐ°ÑÑÐ²Ð¾ÑÐ¸Ð¼Ð¾ÑÑÐ¸ Ð±ÐµÐ»ÐºÐ° L1, Ð¾Ð´Ð½Ð°ÐºÐ¾ ÑÐ¿Ð¾ÑÐ¾Ð±ÑÑÐ²ÑÐµÑ ÐµÐ³Ð¾ Ð½Ð°ÑÐ°Ð±Ð¾ÑÐºÐµ. ÐÐ¿ÑÐ¸Ð¼Ð°Ð»ÑÐ½Ð°Ñ ÐºÐ¾Ð½ÑÑÑÑÐºÑÐ¸Ñ Ð´Ð»Ñ ÑÐ²ÐµÐ»Ð¸ÑÐµÐ½Ð¸Ñ ÑÐ°ÑÑÐ²Ð¾ÑÐ¸Ð¼Ð¾ÑÑÐ¸ ÑÐµÐ»ÐµÐ²Ð¾Ð³Ð¾ Ð±ÐµÐ»ÐºÐ° â ÑÐºÐ¾ÑÐ¾ÑÐµÐ½Ð¸Ðµ Ð³ÐµÐ½Ð° L1 Ð½Ð° 26 Ð°Ð¼Ð¸Ð½Ð¾ÐºÐ¸ÑÐ»Ð¾ÑÐ½ÑÑ Ð¾ÑÑÐ°ÑÐºÐ¾Ð² Ñ N-ÐºÐ¾Ð½ÑÐ° Ð¸ Ð¸ÑÐ¿Ð¾Ð»ÑÐ·Ð¾Ð²Ð°Ð½Ð¸Ðµ ÑÐºÑÐ¿ÑÐµÑÑÐ¸Ð¾Ð½Ð½ÑÑ Ð²ÐµÐºÑÐ¾ÑÐ¾Ð² Ñ ÑÑÐ³Ð°Ð¼Ð¸. ÐÐ°Ð»ÑÐ½ÐµÐ¹ÑÐµÐµ Ð¸ÑÑÐ»ÐµÐ´Ð¾Ð²Ð°Ð½Ð¸Ðµ Ð±ÑÐ´ÐµÑ Ð½Ð°Ð¿ÑÐ°Ð²Ð»ÐµÐ½Ð¾ Ð½Ð° Ð¾ÑÐ¸ÑÑÐºÑ Ð¿Ð¾Ð»ÑÑÐµÐ½Ð½Ð¾Ð³Ð¾ Ð² Ð´Ð°Ð½Ð½Ð¾Ð¹ ÑÐ°Ð±Ð¾ÑÐµ ÑÐ°ÑÑÐ²Ð¾ÑÐ¸Ð¼Ð¾Ð³Ð¾ Ð±ÐµÐ»ÐºÐ° L1 Ð² Ð½Ð°ÑÐ¸Ð²Ð½ÑÑ ÑÑÐ»Ð¾Ð²Ð¸ÑÑ Ni-Ð°ÑÑÐ¸Ð½Ð½Ð¾Ð¹ ÑÑÐ¾Ð¼Ð°ÑÐ¾Ð³ÑÐ°ÑÐ¸Ð¸ Ñ Ð¿Ð¾ÑÐ»ÐµÐ´ÑÑÑÐ¸Ð¼ Ð¿ÑÐ¾Ð²ÐµÐ´ÐµÐ½Ð¸ÐµÐ¼ Ð¸Ð¼Ð¼ÑÐ½Ð¾ÑÐµÑÐ¼ÐµÐ½ÑÐ½Ð¾Ð³Ð¾ Ð°Ð½Ð°Ð»Ð¸Ð·Ð° ÑÑÐ²Ð¾ÑÐ¾ÑÐ¾Ðº ÐºÑÐ¾Ð²Ð¸ Ð²Ð°ÐºÑÐ¸Ð½Ð¸ÑÐ¾Ð²Ð°Ð½Ð½ÑÑ Ð¸ Ð½ÐµÐ²Ð°ÐºÑÐ¸Ð½Ð¸ÑÐ¾Ð²Ð°Ð½Ð½ÑÑ Ð¿Ð°ÑÐ¸ÐµÐ½ÑÐ¾Ð² Ñ Ð¸ÑÐ¿Ð¾Ð»ÑÐ·Ð¾Ð²Ð°Ð½Ð¸ÐµÐ¼ Ð¿Ð¾Ð»ÑÑÐµÐ½Ð½ÑÑ ÑÐµÐºÐ¾Ð¼Ð±Ð¸Ð½Ð°Ð½ÑÐ½ÑÑ Ð±ÐµÐ»ÐºÐ¾Ð².
description: This work is devoted to the production of soluble HPV16 L1 proteins for the further development of an ELISA test system. The tasks of the research: 1. Conduct a literature review on the topic under study. 2. Create genetic constructs with modifications of the L1 gene. 3. Check the effect of shortening the protein L1 gene on its solubility. 4. Create genetic constructs of the modified L1 protein with tags. 5. Check the effect of tags on the solubility of the modified L1 protein. The object of study is the L1 protein of the HPV16 papillomavirus. The subject of the study is the solubility of the L1 protein. The work was carried out on the basis of the Laboratory of Molecular Genetics of Pathogenic Microorganisms of the Pasteur Research Institute of Microorganisms. 3 genetic constructs were collected using the restriction-ligation method with different variants of modification of the L1 gene, as well as 9 genetic constructs with additional tags: MBP, GST, TF and a modified L1 gene. To analyze the efficiency of protein production, as well as to determine the solubility of the protein of interest, the following steps were carried out: cultivation, preparation of fractions and samples for protein electrophoresis in polyacrylamide gel in the presence of sodium dodecyl sulfate according to the Laemmli method. The study revealed that gene shortening does not increase the solubility of the L1 protein, but does contribute to its production, in contrast to the full-length L1 gene. The optimal design for increasing the solubility of the target protein is shortening the L1 gene by 26 amino acid residues from the N-terminus and using an expression vector with tags. Further research will be aimed at purifying the soluble L1 protein obtained in this work under native conditions of Ni-affinity chromatography, followed by enzyme-linked immunosorbent assay of recombinant proteins and blood sera of vaccinated and unvaccinated patients.