استفاده از دو شاتل وکتور درکلون سازی و بیان ژن اینترلوکین 11 در باسیلوس سوبتیلیس
الموضوعات :
آتنا آذرنوش
1
,
مجید مقبلی
2
,
فرشید کفیل زاده
3
,
محمد کارگر
4
,
هوشنگ جمالی
5
1 - گروه میکروبیولوژی دانشگاه آزاد اسلامی جهرم
2 - عضوهیئت علمی /دانشگاه آزاد اسلامی واحد دامغان/دامغان /ایران
3 - دانشگاه آزاد اسلامی، واحد جهرم، گروه میکروبیولوژی
4 - دانشگاه آزاد اسلامی، واحد جهرم، گروه میکروبیولوژی
5 - دانشگاه ازاد اسلامی جهرم
استاد یار
الکلمات المفتاحية: باسیلوس سوبتیلیس, شاتل وکتور, اینترلوکین-11, pHT43, pMR12,
ملخص المقالة :
سابقه و هدف: در بیماران سرطانی که تحت شیمی درمانی قرار می‌گیرند پلاکت خونشان پایین می‌آید اینترلوکین 11، یک سایتوکین افزاینده پلاکت خون است. اینترلوکین 11 نوترکیب انسانی، تنها دارویی است که برای درمان اثرات ناشی از شیمی درمانی تأیید شده است. در این تحقیق از دو شاتل وکتور pHT43 و pMR12 جهت کلون کردن و بیان ژن اینترلوکین 11در باسیلوس سوبتیلیس استفاده شد و میزان بیان در این دو وکتور بررسی گردید.مواد و روش ها: در این مطالعه ژن اینترلوکین 11 به صورت ساختاری بسته با دو جایگاه برشBamHI وXbal با اندازه نهایی bp609 طراحی و سنتز شد. سپس این ژن بر روی شاتل وکتورهای pHT43 و pMR12 کلون و به باکتری باسیلوس سوبتیلیس WB600 انتقال یافت. میزان بیان پروتین نوترکیب اینترلوکین 11 بوسیله معرف بردفورد مورد بررسی قرار گرفت.یافته ها: با روشPCR وجود ژن اینترلوکین 11 را بر روی شاتل وکتورهای pHT43 و pMR12 نشان می‌دهد. میزان بیان توسط باکتری حاملpMR12-int11 پس از 9 ساعت انکوباسیون بیشترین میزان یعنی حدود µg/mL 75 و باکتری حامل pHT43-int11 پس از 4 ساعت، به میزانµg/mL 61/4 بوده است.نتیجه گیری: میزان بیان پروتین نوترکیب اینترلوکین 11 با استفاده از وکتور pMR12 نسبت به وکتور pHT43 بیشتر می‌باشد و تاکنون چنین میزان بیانی برای اینترلوکین 11 گزارش نشده است. باسیلوس سوبتیلیس قادر به بیان و تولید پروتین اینترلوکین 11 می‌باشد و می‌توان از آن به عنوان یک میزبان مناسب برای تولید دارو استفاده نمود.
1994;15(2-1): 9-45.
2. Tanaka.T, Kishimoto.T. The Biology and Medical Implications of Interleukin-6. Cancer
Immunol Res; 2(4) April 2014 chemistry. 1993;268(29) 21527-32.
3. Orazi A, Du X, Yang Z, Kashai M, Williams DA. Interleukin-11 prevents apoptosis and
accelerates recovery of small intestinal mucosa in mice treated with combined chemotherapy
and radiation. Laboratory investigation; a journal of technical methods and pathology. 1996;75
(1): 33-42.
4. Mehler MF, Rozental R, Dougherty M, Spray DC, Kessler JA. Cytokine regulation of neuronal
differentiation of hippocampal progenitor cells. Nature. 1993;362(6415):625.
5. Baumann H, Schendel P. Interleukin-11 regulates the hepatic expression of the same plasma
protein genes as interleukin-6. J. biol. chem. 1991;266(30): 2042-7.
6. Fukuda Y, Sassa S. Effect of interleukin-11 on the levels of mRNAs encoding heme oxygenase
and haptoglobin in human HepG hepatoma cells. Bioche. Bioph. Res. Commun. 1993;193(1):
297- 302.
7. Girasole G, Passeri G, Jilka RL, Manolagas SC. Interleukin-11: a new cytokine critical for
osteoclast development. J. clinic. invest. 1994;93(4): 1516-24.
8. Maier R, Ganu V, Lotz M. Interleukin-11, an inducible cytokine in human articular
chondrocytes and synoviocytes, stimulates the production of the tissue inhibitor of
metalloproteinases. J. biolog.1993 ;15;268(29):21527-32.
9. Peterson RL, Bozza MM, Dorner AJ. Interleukin-11 induces intestinal epithelial cell growth
arrest through effects on retinoblastoma protein phosphorylation. American j. patho. 1994;149
(3) 895-902.
10. Schwertschlag US, Trepicchio WL, Dykstra KH, Keith JC, Turner KJ, Dorner AJ.
Hematopoietic, immunomodulatory and epithelial effects of interleukin-11. Leukemia.
1999;13(9): 1307-15.
11. Kawashima I, Ohsumi J, Mita-Honjo K, Shimoda-Takano K, Ishikawa H, Sakakibara S, et al.
Molecular cloning of cDNA encoding adipogenesis inhibitory factor and identity with
interleukin-11. FEBS letters. 1991;283(2):199-202.
12. Keller DC, Du XX, Srour EF, Hoffman R, Williams DA. Interleukin-11 inhibits adipogenesis
and stimulates myelopoiesis in human long-term marrow cultures. Blood. 1993;82(5):1428-35.
13. Montazeri-Najafabady N, Ghasemi Y, Ali Mobasher M, Ghasemian A, Rasoul-Amini S,
Ebrahimi. Codon Optimization, Cloning and Expression of Interleukin 11 in Two Different E.
coli. Sys.J.Pure App. Microb.2016;7(4):2717-2722.
14. McKinley D, Wu Q, Yang-Feng T, Yang YC. Genomic sequence and chromosomal location
of human interleukin-11 gene (IL11). Genomic.1993; 814(3),13.
15. Shabani D, Kermani SH. Interleukins and tumor tissue death factor. The first international
conference on new findings in biotechnology.2017; https://civilica.com/doc/633058
16. Ye J, Wang Z, Ye D, et al. Increased interleukin-11 levels are correlated with cardiac events in
patients with chronic heart failure. Mediators Inflamm. 2019;15754-10
17. Cook S.A, Schafer S. Hiding in Plain Sight: Interleukin-11 Emerges as a Master Regulator of
Fibrosis, Tissue Integrity, and Stromal Inflammation. Annu. Rev. Med. 2020; 71:263–76
18. Yuchen Li, Qingxin Wu, Yuxin Jin and Qian Yang.Antiviral activity of interleukin-11
as a response to porcine epidemic diarrhea virus infection. Vet. Res .2019; 50:111
19. Widjaja AA, Dong J, Adami E, et al. Redefining interleukin 11 as a regeneration-limiting
hepatotoxin. bioRxiv 830018. https://doi.org/10. 2019; 1101/830018
20. Hirayama F, Katayama N, Neben S, Donaldson D, Nickbarg EB, Clark SC, et al. Synergistic
interaction between interleukin-12 and steel factor in support of proliferation of murine lymphohematopoietic progenitors in culture. Blood.1994;83(1):8-92
21. Walmsley M, Butler DM,Marinova-Mutafchieva L, et al. An anti-inflammatory role for interleukin11 in established murine collagen-induced arthritis. Immunology, 1998; 95(1):31–37.
22. Anguita J, Barthold SW, Samanta S, et al. Selective anti-inflammatory action of interleukin-11
in murine Lyme disease: arthritis decreases while carditis persists. J. Infect. Dis. 1999;179
(3):734–37
23. Elshabrawy HA, Volin MV, Essani AB, et al. IL-11 facilitates a novel connection between
RA joint fibroblasts and endothelial cells. Angiogenesis.2018; 21(2):215–28
24. Kuo D, Ding J, Cohn IS, et al. HBEGF+ macrophages in rheumatoid arthritis induce
fibroblast invasiveness. Sci. Transl. Med. 2019; 11(491): 85-87.
25. Luan C, Zhang HW, Song DG, Xie YG, Feng J, Wang YZ. Expressing antimicrobial peptide
cathelicidin-BF in Bacillus subtilis using SUMO technology. Appl Microbiol Biotechnol.
2014;98(8):3651-8.
26. Rashidi M R, Moghbeli M. Construction of an expression shuttle vector for Escherichia coli
and Bacillus subtilis. J. microbial word. 2013, 3(16), 188-197. [In Persian]
27. Martinez M. A, Dezar C, Baigori M, Sineriz F. Simple method for plasmid mediated
transformation of different Bacillus species. Biotech. Tech.1999; 13(5):337-340.
28. Tepler I, Elias L, Smith JW, et al. A randomized placebo-controlled trial of recombinant
human interleukin- 11 in cancer patients with severe thrombocytopenia due to chemotherapy.
Blood., 1996, (87): 3607- 3614.
29. Hirayama F, Katayama N, Neben S, Donaldson D, Nickbarg EB, Clark SC, et al. Synergistic
interaction between interleukin-12 and steel factor in support of proliferation of murine
lymphohematopoietic progenitors in culture. Blood. 1994; 83(1):8-92
30. Liu N-W, Huang X, Liu S, et al. Elevated BNP caused by recombinant human interleukin-11
treatment in patients with chemotherapy-induced thrombocytopenia. Supp. Care Cancer.
2019;27(11):4293– 98.
31. Miao J, Wang J, Peng S, Tang P, Zou M, Duan J, Zhao C, Ma X. Expression of human
interleukin-11 cDNA in E. coli. 1995;38(10):1202-9.
32. Tan H, Dan G, Gong H, Cao L. Purification and characterization of recombinant truncated
human interleukin-11 expressed as fusion protein in Escherichia coli. Biotech. letters. 2005;
27(13): 905-910.
33. Hosseinian H, Borzamin B. L- asparaginase enzyme gene cloning research and evaluation in
Bacillus subtilis. Cell.Mol. Biotech. News .2014; 4(16), 61-66[In Persian].
34. Sadeghi A, Mahdieh M, Salimi S. Production of Recombinant Human Interleukin-11 (IL-11)
in Transgenic Tobacco (Nicotiana tabacum). J. Plant Biotech.2016;43(4):432-437.
35. Farajnia S, Hasanpour R, Lotfipour F. Cloning and expression of human IL-11 in E. coli .
Pharma.Sci. 2010, 15(4), 353- 359
_||_
1994;15(2-1): 9-45.
2. Tanaka.T, Kishimoto.T. The Biology and Medical Implications of Interleukin-6. Cancer
Immunol Res; 2(4) April 2014 chemistry. 1993;268(29) 21527-32.
3. Orazi A, Du X, Yang Z, Kashai M, Williams DA. Interleukin-11 prevents apoptosis and
accelerates recovery of small intestinal mucosa in mice treated with combined chemotherapy
and radiation. Laboratory investigation; a journal of technical methods and pathology. 1996;75
(1): 33-42.
4. Mehler MF, Rozental R, Dougherty M, Spray DC, Kessler JA. Cytokine regulation of neuronal
differentiation of hippocampal progenitor cells. Nature. 1993;362(6415):625.
5. Baumann H, Schendel P. Interleukin-11 regulates the hepatic expression of the same plasma
protein genes as interleukin-6. J. biol. chem. 1991;266(30): 2042-7.
6. Fukuda Y, Sassa S. Effect of interleukin-11 on the levels of mRNAs encoding heme oxygenase
and haptoglobin in human HepG hepatoma cells. Bioche. Bioph. Res. Commun. 1993;193(1):
297- 302.
7. Girasole G, Passeri G, Jilka RL, Manolagas SC. Interleukin-11: a new cytokine critical for
osteoclast development. J. clinic. invest. 1994;93(4): 1516-24.
8. Maier R, Ganu V, Lotz M. Interleukin-11, an inducible cytokine in human articular
chondrocytes and synoviocytes, stimulates the production of the tissue inhibitor of
metalloproteinases. J. biolog.1993 ;15;268(29):21527-32.
9. Peterson RL, Bozza MM, Dorner AJ. Interleukin-11 induces intestinal epithelial cell growth
arrest through effects on retinoblastoma protein phosphorylation. American j. patho. 1994;149
(3) 895-902.
10. Schwertschlag US, Trepicchio WL, Dykstra KH, Keith JC, Turner KJ, Dorner AJ.
Hematopoietic, immunomodulatory and epithelial effects of interleukin-11. Leukemia.
1999;13(9): 1307-15.
11. Kawashima I, Ohsumi J, Mita-Honjo K, Shimoda-Takano K, Ishikawa H, Sakakibara S, et al.
Molecular cloning of cDNA encoding adipogenesis inhibitory factor and identity with
interleukin-11. FEBS letters. 1991;283(2):199-202.
12. Keller DC, Du XX, Srour EF, Hoffman R, Williams DA. Interleukin-11 inhibits adipogenesis
and stimulates myelopoiesis in human long-term marrow cultures. Blood. 1993;82(5):1428-35.
13. Montazeri-Najafabady N, Ghasemi Y, Ali Mobasher M, Ghasemian A, Rasoul-Amini S,
Ebrahimi. Codon Optimization, Cloning and Expression of Interleukin 11 in Two Different E.
coli. Sys.J.Pure App. Microb.2016;7(4):2717-2722.
14. McKinley D, Wu Q, Yang-Feng T, Yang YC. Genomic sequence and chromosomal location
of human interleukin-11 gene (IL11). Genomic.1993; 814(3),13.
15. Shabani D, Kermani SH. Interleukins and tumor tissue death factor. The first international
conference on new findings in biotechnology.2017; https://civilica.com/doc/633058
16. Ye J, Wang Z, Ye D, et al. Increased interleukin-11 levels are correlated with cardiac events in
patients with chronic heart failure. Mediators Inflamm. 2019;15754-10
17. Cook S.A, Schafer S. Hiding in Plain Sight: Interleukin-11 Emerges as a Master Regulator of
Fibrosis, Tissue Integrity, and Stromal Inflammation. Annu. Rev. Med. 2020; 71:263–76
18. Yuchen Li, Qingxin Wu, Yuxin Jin and Qian Yang.Antiviral activity of interleukin-11
as a response to porcine epidemic diarrhea virus infection. Vet. Res .2019; 50:111
19. Widjaja AA, Dong J, Adami E, et al. Redefining interleukin 11 as a regeneration-limiting
hepatotoxin. bioRxiv 830018. https://doi.org/10. 2019; 1101/830018
20. Hirayama F, Katayama N, Neben S, Donaldson D, Nickbarg EB, Clark SC, et al. Synergistic
interaction between interleukin-12 and steel factor in support of proliferation of murine lymphohematopoietic progenitors in culture. Blood.1994;83(1):8-92
21. Walmsley M, Butler DM,Marinova-Mutafchieva L, et al. An anti-inflammatory role for interleukin11 in established murine collagen-induced arthritis. Immunology, 1998; 95(1):31–37.
22. Anguita J, Barthold SW, Samanta S, et al. Selective anti-inflammatory action of interleukin-11
in murine Lyme disease: arthritis decreases while carditis persists. J. Infect. Dis. 1999;179
(3):734–37
23. Elshabrawy HA, Volin MV, Essani AB, et al. IL-11 facilitates a novel connection between
RA joint fibroblasts and endothelial cells. Angiogenesis.2018; 21(2):215–28
24. Kuo D, Ding J, Cohn IS, et al. HBEGF+ macrophages in rheumatoid arthritis induce
fibroblast invasiveness. Sci. Transl. Med. 2019; 11(491): 85-87.
25. Luan C, Zhang HW, Song DG, Xie YG, Feng J, Wang YZ. Expressing antimicrobial peptide
cathelicidin-BF in Bacillus subtilis using SUMO technology. Appl Microbiol Biotechnol.
2014;98(8):3651-8.
26. Rashidi M R, Moghbeli M. Construction of an expression shuttle vector for Escherichia coli
and Bacillus subtilis. J. microbial word. 2013, 3(16), 188-197. [In Persian]
27. Martinez M. A, Dezar C, Baigori M, Sineriz F. Simple method for plasmid mediated
transformation of different Bacillus species. Biotech. Tech.1999; 13(5):337-340.
28. Tepler I, Elias L, Smith JW, et al. A randomized placebo-controlled trial of recombinant
human interleukin- 11 in cancer patients with severe thrombocytopenia due to chemotherapy.
Blood., 1996, (87): 3607- 3614.
29. Hirayama F, Katayama N, Neben S, Donaldson D, Nickbarg EB, Clark SC, et al. Synergistic
interaction between interleukin-12 and steel factor in support of proliferation of murine
lymphohematopoietic progenitors in culture. Blood. 1994; 83(1):8-92
30. Liu N-W, Huang X, Liu S, et al. Elevated BNP caused by recombinant human interleukin-11
treatment in patients with chemotherapy-induced thrombocytopenia. Supp. Care Cancer.
2019;27(11):4293– 98.
31. Miao J, Wang J, Peng S, Tang P, Zou M, Duan J, Zhao C, Ma X. Expression of human
interleukin-11 cDNA in E. coli. 1995;38(10):1202-9.
32. Tan H, Dan G, Gong H, Cao L. Purification and characterization of recombinant truncated
human interleukin-11 expressed as fusion protein in Escherichia coli. Biotech. letters. 2005;
27(13): 905-910.
33. Hosseinian H, Borzamin B. L- asparaginase enzyme gene cloning research and evaluation in
Bacillus subtilis. Cell.Mol. Biotech. News .2014; 4(16), 61-66[In Persian].
34. Sadeghi A, Mahdieh M, Salimi S. Production of Recombinant Human Interleukin-11 (IL-11)
in Transgenic Tobacco (Nicotiana tabacum). J. Plant Biotech.2016;43(4):432-437.
35. Farajnia S, Hasanpour R, Lotfipour F. Cloning and expression of human IL-11 in E. coli .
Pharma.Sci. 2010, 15(4), 353- 359
