Summary
Antisense RNA targeted against the major capsid protein (MCP) of Lactococcus lactis subsp. cremoris bacteriophage F4-1 reduced bacteriophage replication by up to 50%. The region containing the mcp gene was oriented to transcribe the antisense strand using a L. lactis subsp. cremoris Wg2 promoter. The size of the mcp insert transcribed affected the level of bacteriophage inhibition and the greatest level of inhibition was achieved using a 301-bp fragment from the 5′ end of the mcp. Antisense mcp RNA constructs were stable and did not alter the endogenous plasmid profile in the host, L. lactis subsp. cremoris F4-1. There were, however, some adverse effects on the host during the stationary phase as exhibited by a decline in cell density.
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References
Alatossava T, Klaenhammer TR (1991) Molecular characterization of three small isometric-headed bacteriophages which vary in their sensitivity to the lactococcal phage resistance plasmid pTR2030. Appl Environ Microbiol 57:1346–1353
Andersen J, Forst SA, Zhao K, Inouye M, Delihas N (1989) The function of micF RNA. J Biol Chem 264:17961–17979
Anderson DG, McKay LL (1983) Simple and rapid method for isolating large plasmid DNA from lactic streptococci. Appl Environ Microbiol 46:549–552
Boizet B, Lahbib-Mansais Y, Dupont L, Ritzenthaler P, Mata M (1991) Cloning, expression and sequence analysis of an endolysin-encoding gene of Lactobacillus bulgaricus bacteriophage mv1. Gene 94:61–67
Chung DK, Kim JK, Batt CA (1991) Cloning and nucleotide sequence of the major capsid protein from Lactococcus lactis ssp. cremoris bacteriophage F4–1. Gene 101:121–125
Coleman J, Hirasuma A, Inokuchi Y, Green PJ, Inouye M (1985) A novel immune system against bacteriophage infection using complementary RNA (micRNA). Nature 315:601
Ecker J, Davis R (1986) Inhibition of gene expression in plant cells by expression of antisense RNA. Proc Natl Acad Sci USA 83:5372–5376
Hanahan D (1983) Studies on transformation of Escherichia coli with plasmids. J Mol Biol 166:557–580
Hill C, Miller LA, Klaenhammer TR (1990a) Nucleotide sequence and distribution of the pTR2030 resistance determinant (hsp) which aborts bacteriophage infection in lactococci. Appl Environ Microbiol 56:2255–2258
Hill C, Miller LA, Klaenhammer TR (1990b) Cloning, expression, and sequence determination of a bacteriophage fragment encoding bacteriophage resistance in Lactococcus lactis. J Bacteriol 172:6419–6426
Hill C, Miller LA, Klaenhammer TR (1991) In vivo genetic exchange of a functional domain from a type II A methylase between lactococcal plasmid pTR2030 and a virulent bacteriophage. J Bacteriol 173:4363–4370
Hirashima A, Sawaki S, Inokuchi Y, Inouye M (1986) Engineering of the mRNA-interfering complementary RNA immune system against viral infection. Proc Natl Acad Sci USA 3:7726–7730
Innis MA, Gelfand DH (1990) Optimizations of PCRs. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ (eds) PCR protocols: a guide to methods and applications. Academic Press, San Diego, Calif., pp 3–12
Inouye M (1988) Antisense RNA: its functions and applications in gene regulation — a review. Gene 72:25–34
Izant J, Weintraub H (1984) Inhibition of thymidine kinase gene expression by anti-sense RNA: a molecular approach to genetic analysis. Cell 36:1007–1115
Jarvis AW (1989) Bacteriophages of lactic acid bacteria. J Dairy Sci 72:3406–3428
Kim JH, Batt CA (1991a) Molecular characterization of a Lactococcus lactis bacteriophage F4–1. Food Microbiol 8:15–26
Kim JH, Batt CA (1991b) Nucleotide sequence and deletion analysis of a gene coding for a structural protein of Lactococcus lactis bacteriophage F4–1. Food Microbiol 8:27–36
Kim SG, Batt CA (1991c) Identifucation of a nucleotide sequence conserved in Lactococcus lactis bacteriophages. Gene 98:95–100
Kim SG, Batt CA (1991d) Antisense mRNA mediated bacteriophage resistance in Lactococcus lactis. Appl Environ Microbiol 57:1109–1113
Kok J, Vossen JMBM van der, Venema G (1984) Construction of plasmid cloning vectors for lactic streptococci which also replicate in Bacillus subtilis and Escherichia coli. Appl Environ Microbiol 48:726–731
Lakshmidevi G, Davidson BE, Hillier AJ (1988) Circular permutation of the genome of a temperate bacteriophage from Streptococcus lactis. Appl Environ Microbiol 54:1039–1045
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Press, Cold Spring Harbor, N. Y.
Marshall RJ, Berridge NJ (1976) Selection and some properties of phage-resistant starter for cheese-making. J Dairy Res 43:449–458
Mizuno T, Chou M-Y, Inouye M (1984) A unique mechanism regulating gene expression: translational inhibition by a complementary RNA transcript (micRNA) Proc Natl Acad Sci USA 81:1966–1970
Pestka S, Daugherty BL, Jung V, Hotta K, Pestka RK (1984) Anti-mRNA: specific inhibition of translation of single mRNA molecules. Proc Natl Acad Sci USA 81:7525–7528
Scharf SJ (1990) Cloning with PCR. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ (eds) PCR protocols: a guide to methods and applications. Academic Press, San Diego, Calif., pp 84–91
Shearman C, Underwood H, Jury K, Gasson M (1989) Cloning and DNA sequence analysis of a Lactococcus bacteriophage lysin gene. Mol Gen Genet 218:214–221
Stephenson ML, Zamecnik PC (1978) Inhibition of Rous sarcoma viral RNA translation by a specific oligodeoxyribonucleotides. Proc Natl Acad Sci USA 75:285–288
Terzaghi BE, Sandine WE (1975) Improved medium for lactic streptococci and their bacteriophages. Appl Microbiol 29:807–817
Thunell R, Sandine W, Bodyfelt F (1981) Phage-insensitive, multiple-strain starter approach to cheddar cheese. J Dairy Sci 64:2270–2277
Vieira J, Messing J (1982) The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene 19:259–268
Vossen JMBM van der, Lelie D, van der, Venema G (1987) Isolation and characterization of Streptococcus cremoris Wg2-specific promoters. Appl Environ Microbiol 53:2452–2457
Whitehead HR, Cox GA (1936) Bacteriophage phenomena in cultures of lactic streptococci. J Dairy Res 7:55–62
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Kyun Chung, D., Kyun Chung, S. & Batt, C.A. Antisense RNA directed against the major capsid protein of Lactococcus lactis subsp. cremoris bacteriophage 4-1 confers partial resistance to the host. Appl Microbiol Biotechnol 37, 79–83 (1992). https://doi.org/10.1007/BF00174207
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DOI: https://doi.org/10.1007/BF00174207