Removal of the selective marker gene in the fusion gene expression vector of plant anti-caries vaccine

doi:10.12307/2023.767

Chinese Journal of Tissue Engineering Research ›› 2024, Vol. 28 ›› Issue (1): 7-11.doi: 10.12307/2023.767

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Removal of the selective marker gene in the fusion gene expression vector of plant anti-caries vaccine

Lang Yaoling1, 2, Wang Qian1, Chen Bin1, Bai Guohui1, Guan Xiaoyan1, Liu Jianguo1

1Guizhou Provincial Key Laboratory of Oral Disease Research in Colleges and Universities and Zunyi Key Laboratory of Oral Disease Research, School of Stomatology, Zunyi Medical University, Zunyi 563000, Guizhou Province, China; 2Guiyang Stomatological Hospital, Guiyang 550002, Guizhou Province, China

Received:2022-11-03 Accepted:2022-12-24 Online:2024-01-08 Published:2023-06-28
Contact: Liu Jianguo, MD, Professor, Chief physician, Doctoral supervisor, Guizhou Provincial Key Laboratory of Oral Disease Research in Colleges and Universities and Zunyi Key Laboratory of Oral Disease Research, School of Stomatology, Zunyi Medical University, Zunyi 563000, Guizhou Province, China
About author:Lang Yaoling, Master, Attending physician, Guizhou Provincial Key Laboratory of Oral Disease Research in Colleges and Universities and Zunyi Key Laboratory of Oral Disease Research, School of Stomatology, Zunyi Medical University, Zunyi 563000, Guizhou Province, China; Guiyang Stomatological Hospital, Guiyang 550002, Guizhou Province, China
Supported by:
the National Natural Science Foundation of China, No. 81260164 (to LJG); The Sixth Batch of Talent Bases in Guizhou Province of the Organization Department of Guizhou Provincial Party Committee, No. RCJD2019-9 (to LJG); The First Batch of Zunyi Municipal Talent Bases, No. Zunyi KHSZ[2017]20 (to LJG); Young Scientific and Technological Talent Growth Project of the Department of Education of Guizhou Province, No. QJHKYZ[2022]276 (to CB); Science and Technology Fund Project of Guizhou Provincial Health Commission, No. gzwkj2021-351 (to CB)

Abstract

Abstract: BACKGROUND: In consideration of the food safety and ecological safety of transgenic plants, the retention of marker genes is the primary safety issue affecting transgenic plants.
OBJECTIVE: Based on the principle of immune caries prevention, our research team successfully constructed the plant anti-caries vaccine fusion gene expression vector pCAMBIA-E8-APB-DOCK8 for these two caries causing virulence factors surface protein and glucosyltransferase, which provides a basis for the research and development of transgenic plant vaccine.
METHODS: In this study, the selective marker genes Km and GUS in the plant caries vaccine fusion gene expression vector pCAMBIA-E8-APB-DOCK8 were removed by DNA recombination technology through a series of steps such as DNA fragment separation, connection, transformation, clone detection, and sequencing.
RESULTS AND CONCLUSION: The efficiency of marker gene removal was 99%. This study has laid a good experimental foundation for the safe production of transgenic plant vaccine against dental caries, and also provided ideas for the construction of other plant vaccine vectors.

Key words: plant vaccine, immune anti-caries, expression vector, marker gene, transgenic safety, E8 promoter

CLC Number:

Lang Yaoling, Wang Qian, Chen Bin, Bai Guohui, Guan Xiaoyan, Liu Jianguo. Removal of the selective marker gene in the fusion gene expression vector of plant anti-caries vaccine[J]. Chinese Journal of Tissue Engineering Research, 2024, 28(1): 7-11.

Figures/Tables 4

References

[1] 王兴.第四次全国口腔健康流行病学调查报告[M].北京:人民卫生出版社, 2018.
[2] YUKTA N, NIKHIL M, RUTUJA R, et al. Pit and Fissure Sealants. J Res Med Dent Sci. 2022;10(1):52-56.
[3] ANETA M, ALINA-MARIA H, MIHAELA-RODICA P, et al. Review of Professionally Applied Fluorides for Preventing Dental Caries in Children and Adolescents. Appl Sci. 2022;12(1):1054.
[4] ARSLAN S, KAPLAN MH. The Effect of Resin Infiltration on the Progression of Proximal Caries Lesions: A Randomized Clinical Trial. Med Princ Pract. 2020;29(3): 238-243.
[5] VALENTINA L, ANTONIO M, LUCA R, et al. Ozone Therapy for Early Childhood Caries (ECC) Treatment: An In Vivo Prospective Study. Appl Sci. 2022;12(1):1964.
[6] HOSHINO T, FUJIWARA T. The findings of glucosyltransferase enzymes derived from oral streptococci. Jpn Dent Sci Rev. 2022;58:328-335.
[7] CHERUKURI G, VEERAMACHANENI C, RAO GV, et al. Insight into status of dental caries vaccination: A review. J Conserv Dent. 2020;23(6):544-549.
[8] SALYAEV RK, REKOSLAVSKAYA NI, STOLBIKOV AS, et al. The study of immunogenicity of the antigenic protein of high risk oncogenic type of the human papillomavirus HPV16 L1 produced in the plant expression system on the base of transgenic tomato. Dokl Biochem Biophys. 2017;474(1):186-188.
[9] 孙宇辰.基因工程疫苗的研究与应用[J].生物化工,2018,4(6):152-158.
[10] 田源,刘建国,白国辉,等.表达嵌合体蛋白 PAcP/CTB 的转基因番茄口服免疫大白兔的实验研究[J].口腔医学研究,2013,29(3):201-204.
[11] 顾瑜.转基因番茄可食防龋疫苗免疫大鼠的实验研究[J].重庆医学,2015, 44(23):3169-3175.
[12] 顾瑜.表达嵌合体蛋白PAcP/CTB的转基因番茄免疫BABL/c小鼠的实验研究[J].牙体牙髓牙周病学杂志,2014,24(2):72-75.
[13] 李虎,管晓燕,李敏,等.防龋基因疫苗pVAX1-GbpA/GBD缓释温敏型水凝胶不同途径免疫兔的效果研究[J].儿童龋病研究,2022,38(7):640-646.
[14] 关薇薇,顾瑜,管晓燕,等.表达嵌合蛋白PAcA/CTB 转基因番茄疫苗可通过胃肠道吸收的方式免疫大鼠[J].中国组织工程研究,2021,25(11):1712-1716.
[15] 李虎,管晓燕,李敏,等.水凝胶负载pVAX1-SpaP/P防龋DNA疫苗以4种途径产生免疫效应[J].中国组织工程研究,2022,26(27):4340-4345.
[16] 关薇薇,顾瑜,管晓燕,等.转基因番茄防龋疫苗中外源目的基因鉴定及表达检测[J].中国组织工程研究,2022,26(2):171-175.
[17] YONGRUI H, LANZHEN X, AIHONG P, et al. Production of marker-free transgenic plants from mature tissues of navel orange using a Cre/loxP site-recombination system. Hortic Plant J. 2022;10(1):1-10.
[18] SINGH R, KAUR N, PRABA UP, et al. A Prospective Review on Selectable Marker-Free Genome Engineered Rice: Past, Present and Future Scientific Realm. Front Genet. 2022;13:882836.
[19] 范繁.E8-APB-DOCK8融合基因表达质粒转化番茄的研究[D].遵义:遵义医学院,2015.
[20] GHANEM S. Cloning of the nptII gene of Escherichia coli and construction of a recombinant strain harboring functional recA and nptII antibiotic resistance. Genet Mol Res. 2011;10(3):1445-1454.
[21] HU H, YU F. A CRISPR/Cas9-Based System with Controllable Auto-Excision Feature Serving Cisgenic Plant Breeding and Beyond. Int J Mol Sci. 2022;23(10):5597.
[22] ARSALAN R, MOHAMMAD F, SAEID M, et al. In planta removal of nptII selectable marker gene from transgenic tobacco plants using CRISPR/Cas9 system. Plant Gene. 2021;26(1):100288.
[23] JACOB S,VELUTHAMBI K. Generation of selection marker-free transgenic plants by cotransformation of a cointegrate vector T-DNA and a binary vector T-DNA in one Agrobacterium tumefaciens strain. Plant Science. 2002;163:801-806.
[24] SMULIAN AG, GIBBONS RS, DEMLAND JA, et al. Expression of hygromycin phosphotransferase alters virulence of Histoplasma capsulatum. Eukaryot Cell. 2007;6(11):2066-2071.
[25] FUCHS RL, REAM JE, HAMMOND BG, et al. Salety assessment of the neomycin photransferase-Ⅱ (NPTⅡ) protein. Biotechnology. 1993;11(13):1543-1547.
[26] MIKI B, MCHUGH S. Selectable marker genes in transgenic plants: applications, alternatives and biosafety. J Biotechnol. 2004;107(3):193-232.
[27] KAI GY, ZHANG L, ZHANG HY, et al. Marker-free:a novel tendency of transgenic plants. Acta Botanica Sinica. 2002;44(8):883-888.
[28] FISCHHOFF DA, BOWDISH KS, PERLAK F, et al. Insect tolerant transgenic tomato plants. Biotechnology. 1987;5(8):807-813.
[29] XUE B, GONSALVES C, PROVVIDENTI R, et al. Development of transgenic tomato expressing a high-level of resistance to cucumber mosaic-virus strains of subgroup-Ⅰand subgroup-Ⅱ. Plant Disease. 1994;78(11):1038-1041.
[30] KUNIK T, SALOMON R, ZAMIR D, et al. Transgenic tomato plants expressing the tomato yellow leaf curl virus capsid protein are resistant to the virus. Biotechnology. 1994;12(5):500-504.
[31] WHITHAM S, MCCORMICK S, BAKER B. The N gene of tobacco confers resistance to tobacco mosaic virus in transgenic tomato. Proc Natl Acad Sci U S A. 1996; 93(16):8776-8781.
[32] THOMZIK JE, STENZEL K, STOCKER R, et al. Synthesis of a grapevine phytoalexin in transgenic tomatoes (Lycopersicon esculentum Mill.) conditionsresistance against Phytophthora infestans. Physiol Mol Plant Pathol. 1997;51(4):265-278.
[33] SANDHU JS, KRASNYANSKI SF, DOMIER LL, et al. Oral immunization of mice with transgenic tomato fruit expressing respiratory syncytial virus-F protein induces a systemic immune response. Transgenic Res. 2000;9(2):127-135.
[34] JANI D, MEENA LS, RIZWAN-UL-HAQ QM, et al. Expression of cholera toxin B subunit in transgenic tomato plants. Transgenic Res. 2002;11(5):447-454.
[35] JIANG XL, HE ZM, PENG ZQ, et al. Cholera toxin B protein in transgenic tomato fruit induces systemic immune response in mice. Transgenic Res. 2007;16(2):169-175.
[36] SHCHELKUNOV SN, SALYAEV RK, REKOSLAVSKAYA NI, et al. The obtaining of transgenic tomato plant producing chimerical proteins TBI-HBsAg. Dokl Biochem Biophys. 2004;396:139-142.
[37] CHEN HF, CHANG MH, CHIANG BL, et al. Oral immunization of mice using transgenic tomato fruit expressing VP1 protein from enterovirus 71. Vaccine. 2006;24(15):2944-2951.
[38] MCGARVEY PB, HAMMOND J, DIENELT MM, et al. Expression of the rabies virus glycoprotein in transgenic tomatoes. Biotechnology (N Y). 1995;13(13):1484-1487.
[39] HU W, ZHOU T, WANG P, et al. Development of Whole-Genome Agarose-Resolvable LInDel Markers in Rice. Rice (N Y). 2020;13(1):1.

Removal of the selective marker gene in the fusion gene expression vector of plant anti-caries vaccine

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