Molecular and Biological Characterization of a New Isolate of Guinea Pig Cytomegalovirus
Abstract
:1. Introduction
2. Results and Discussion
2.1. Isolation of the CIDMTR Strain
2.2. Morphological Analyses by EM
2.3. Sequencing and Sequence Analysis
GPCMV Open Reading Frames (ORFs): CIDMTR Strain | |||||
---|---|---|---|---|---|
ORF | Strand | Begin | End | Codons | Notes |
gp1 | C | 12,464 | 12,769 | 101 | GPCMV MIP 1α; CC chemokine homolog |
gp2 | 14,840 | 15,685 | 281 | Homology to MCMV M69a | |
gp3 | C | 17,197 | 19,563 | 788 | Homology to THV T5b; US22 superfamily |
gp4 | C | 20,829 | 21,152 | 107 | Homology to RCMV r136d |
gp5 | C | 26,721 | 27,818 | 365 | Homology to MCMV m32a |
GP23 | C | 33,299 | 34,501 | 400 | UL23 homolog; US22 gene superfamily |
GP24 | C | 34,739 | 35,956 | 405 | UL24 homolog; US22 superfamily |
GP25 | 36,542 | 38,194 | 550 | UL25 homolog; tegument protein | |
GP26 | C | 38,360 | 39,043 | 227 | UL26 homolog |
GP27 | C | 39,166 | 41,211 | 681 | UL27 homolog |
GP28 | C | 41,311 | 42,378 | 355 | UL28 homolog; US22 superfamily |
GP28.1 | C | 42,866 | 44,287 | 473 | UL28 homolog; US22 superfamily |
GP29 | C | 44,653 | 46,623 | 656 | UL29 homolog; US22 superfamily |
gp29.1 | C | 47,247 | 47,861 | 204 | US22 superfamily |
GP30 | C | 49,082 | 50,800 | 572 | UL30 homolog |
GP31 | 51,094 | 52,572 | 492 | UL31 homolog | |
GP32 | C | 52,665 | 54,365 | 566 | UL32 homolog |
GP33 | 54,585 | 55,868 | 427 | UL33 homolog; 7-TMR GPCR superfamily | |
GP34 | 56,221 | 57,804 | 527 | UL34 homolog | |
GP35 | 58,008 | 59,666 | 552 | UL35 homolog | |
GP37 | C | 59,788 | 60,711 | 307 | UL37 homolog |
GP38 | C | 61,068 | 61,988 | 306 | UL38 homolog |
gp38.1 | C | 62,705 | 63,262 | 185 | Positional homolog of HCMV UL40 |
gp38.2 | C | 63,620 | 64,933 | 437 | Positional homolog of HCMV UL41a |
gp38.3 | C | 65,624 | 66,478 | 284 | Positional homolog of HCMV UL42 |
gp38.4 | C | 66,997 | 67,362 | 121 | Homology to RCMV r42d |
GP43 | C | 67,951 | 68,964 | 337 | UL43 homolog |
GP44 | C | 68,952 | 70,175 | 407 | UL44 homolog |
GP45 | C | 70,887 | 73,673 | 928 | UL45 homolog |
GP46 | C | 73,779 | 74,867 | 362 | UL46 homolog |
GP47 | 74,674 | 77,787 | 1037 | UL47 homolog | |
GP48 | 77,784 | 84,155 | 2123 | UL48 homolog | |
GP48.2 | C | 84,238 | 84,468 | 76 | UL48a homolog |
GP49 | C | 84,479 | 86,119 | 546 | UL49 homolog |
GP50 | C | 86,088 | 87,158 | 356 | UL50 homolog |
GP51 | C | 87,281 | 87,580 | 99 | UL51 homolog; terminase subunit |
GP52 | 87,900 | 89,480 | 526 | UL52 homolog | |
GP53 | 89,473 | 90,459 | 328 | UL53 homolog | |
gp53.1 | C | 90,196 | 90,537 | 113 | Homology to RhCMV rh86; YP_068179.1 |
GP54 | C | 90,551 | 93,904 | 1117 | UL54 homolog; DNA polymerase |
GP55 | C | 93,947 | 96,652 | 901 | UL55 homolog; glycoprotein B |
GP56 | C | 96,549 | 98,816 | 755 | UL56 homolog; terminase subunit |
GP57 | C | 98,967 | 102,650 | 1227 | UL57 homolog |
gp57.1 | C | 104,411 | 104,926 | 171 | Homolog to RCMV r23.1d |
GP69 | C | 108,260 | 111,430 | 1056 | UL69 homolog |
GP70 | C | 112,136 | 115,339 | 1067 | UL70 homolog; helicase-primase |
GP71 | 115,338 | 116,114 | 258 | UL71 homolog | |
GP72 | C | 116,277 | 117,350 | 357 | UL72 homolog; dUTPase |
GP73 | 117,432 | 117,833 | 133 | UL73 homolog; glycoprotein N | |
GP73.5ex1 | 117,860 | 117,869 | 3 | ||
GP73.5ex2 | 119,005 | 119,198 | 64 | ||
GP74 | C | 117,780 | 118,904 | 374 | UL74 homolog; glycoprotein O |
GP75 | C | 119,349 | 121,529 | 726 | UL75 homolog; glycoprotein H |
GP76 | 121,694 | 122,533 | 279 | UL76 homolog | |
GP77 | 122,247 | 124,091 | 614 | UL77 homolog | |
GP78 | 124,473 | 125,717 | 414 | UL78 homolog; 7-TMR GPCR superfamily | |
GP79 | C | 125,914 | 126,861 | 315 | UL79 homolog |
GP80 | 126,722 | 129,031 | 769 | UL80 homolog; CMV protease | |
GP80.5 | 127,610 | 129,031 | 473 | UL80.5 | |
GP82 | C | 129,324 | 130,889 | 521 | UL82 homolog; pp71 |
GP83 | C | 131,109 | 132,809 | 566 | UL83 homolog; pp65 |
GP84 | C | 133,051 | 134,487 | 478 | UL84 homolog |
GP85 | C | 134,785 | 135,696 | 303 | UL85 homolog |
GP86 | C | 135,977 | 140,026 | 1349 | UL86 homolog |
GP87 | 140,407 | 143,328 | 973 | UL87 homolog | |
GP88 | 143,231 | 144,499 | 422 | UL88 homolog | |
GP89ex2 | C | 144,545 | 145,675 | 376 | UL89 homolog; terminase subunit, exon 2 |
GP91 | 146,102 | 146,365 | 87 | UL91 homolog | |
GP92 | 146,362 | 146,991 | 209 | UL92 homolog | |
GP93 | 146,957 | 148,732 | 591 | UL93 homolog | |
GP94 | 148,644 | 149,681 | 345 | UL94 homolog | |
GP89ex1 | C | 150,032 | 150,913 | 291 | UL89 homolog; terminase subunit, exon 1 |
GP95 | 150,967 | 152,235 | 422 | UL95 homolog | |
GP96 | 152,468 | 152,830 | 120 | UL96 homolog | |
GP97 | 152,910 | 154,727 | 605 | UL97 homolog; protein kinase | |
GP98 | 154,747 | 156,531 | 594 | UL98 homolog; alkaline nuclease | |
GP99 | 156,444 | 156,965 | 173 | UL99 homolog; pp28 | |
gp99.1 | 157,155 | 157,769 | 204 | Homology to RCMV r4d | |
GP100 | C | 157,278 | 158,327 | 349 | UL100 homolog; glycoprotein M |
GP102 | 158,657 | 160,942 | 761 | UL102 homolog | |
GP103 | C | 161,127 | 161,852 | 241 | UL103 homolog |
GP104 | C | 161,815 | 163,908 | 697 | UL104 homolog; portal |
GP105 | 163,748 | 166,531 | 927 | UL105 homolog; helicase-primase | |
GP112ex1 | 176,745 | 177,498 | 315 | UL112 homolog; replication accessory, ex 1 | |
GP112ex2 | 177,606 | 177,782 | UL112 homolog; replication accessory, ex 2 | ||
GP112ex3 | 178,115 | 178,131 | UL122 homolog; replication accessory, ex 3 | ||
GP114 | C | 179,126 | 179,920 | 264 | UL114 homolog; uracil glycosylase |
GP115 | C | 179,986 | 180,762 | 258 | UL115 homolog; glycoprotein L |
GP116 | C | 180,755 | 181,654 | 299 | Homology to THV t116b; Fc receptor//Ig |
GP117 | C | 181,877 | 183,262 | 461 | UL117 homolog |
gp119.1 | C | 184,418 | 185,167 | 249 | Similar to MCMV in ACE95619.1 |
GP121.2 | C | 185,160 | 185,834 | 224 | Betaherpesvirus B7D8, accession AFK83957 |
GP121.4 | C | 186,299 | 187,174 | 291 | UL121 homolog; Tupaia t121.4, NP_116476 |
GP122ex3 | C | 187,993 | 189,677 | 677 | UL122 homolog; HCMV IE2 |
GP122ex2 | C | 191,079 | 191,311 | ||
§GP122ex1 | C | 191,403 | 191,518 | ||
GP123ex3 | C | 189,907 | 190,985 | 475 | UL123 homolog; HCMV IE1 |
GP123ex2 | C | 191,079 | 191,311 | ||
§GP123ex1 | C | 191,403 | 191,518 | ||
GP128 | 195,400 | 196,455 | 351 | Similar to Bat HSV B126; US22 Family | |
gp130 | 196,655 | 196,999 | 114 | ||
GP129ex3 | C | 196,432 | 196,690 | 178 | Homolog of HCMV UL128 |
GP129ex2 | 196,768 | 196,890 | |||
GP129ex1 | C | 196,974 | 197,128 | ||
GP131ex2 | C | 197,133 | 197,469 | 191 | Homolog of HCMV UL130 |
GP131ex1 | C | 197,550 | 197,788 | ||
GP133 | C | 197,788 | 198,174 | 128 | Homolog of HCMV UL131 |
GP134 | C | 198,268 | 198,951 | 227 | |
gp138.2 | C | 199,367 | 200,875 | 502 | |
gp138.3 | C | 201,090 | 202,607 | 505 | |
gp139 | C | 202,706 | 204,793 | 695 | THV T5; US22 superfamily |
gp140 | 204,522 | 204,929 | 135 | Homology to CCMV UL132 | |
gp141 | C | 205,053 | 206,660 | 535 | HCMV US23; US22 superfamily |
gp142 | C | 206,928 | 208,622 | 564 | HCMV US24; US22 superfamily |
gp143 | C | 208,875 | 210,866 | 663 | THV T5; US22 superfamily |
gp144 | C | 211,109 | 213,403 | 764 | US26; US22 gene superfamily |
gp145 | C | 213,677 | 215,575 | 632 | HCMV IRS1/TRS1; US22 superfamily |
gp146 | C | 215,932 | 217,914 | 660 | HCMV IRS1/TRS1; US22 superfamily |
gp147.1 | C | 221,739 | 222,935 | 398 | MHC class I homolog |
gp147 | C | 223,124 | 224,218 | 364 | MHC class I homolog |
gp148 | C | 225,379 | 226,560 | 393 | MHC class I homolog |
gp149 | C | 228,236 | 230,209 | 657 | MHC class I homolog |
gp149.1 | 230,163 | 230,465 | 100 | Unique ORF sequence in CIDMTR |
2.4. Sequence Characterization of DNA from Original SG Homogenate in Region of IE1/2 Start Codon
2.5. PCR Confirmation of GPCMV-CIDMTR Genome Structure
Primer Name | Primer Sequence |
---|---|
Mismatch Region 1 F1 | 5'-GTGAGACGTAAGAATAGCTTGC-3' |
Mismatch Region 1 F2 | 5'-GATCCTTAGACTCTATCACGG-3' |
Mismatch Region 1 R1 | 5'-GTGTTGTCACAATTGGCACATG-3' |
Mismatch Region 1 R2 | 5'-ACATGGTCACGACAGAATC-3' |
Mismatch Region 2 F1 | 5'-GTGGACAGGATCCCCAAATT-3' |
Mismatch Region 2 F2 | 5'-CCAAATTTCTGTCGTCGGCG-3' |
Mismatch Region 2 R1 | 5'-TGTTTCCGTGTCTGTCTCCGT-3' |
Mismatch Region 2 R2 | 5'-GTCTTAGCCCGAGACCTTC-3' |
2.6. Infection In Vivo and Development of a Real-Time PCR Assay for Detection of CIDMTR DNA
3. Experimental Section
3.1. Cells, Virus and DNA Preparation
3.2. Transmission Electron Microscopy
3.3. Deep Sequencing and Sequence Analyses
3.4. RT-PCR and RACE Analyses
3.5. GPCMV-CIDMTR Strain PCR Assay
3.6. Animal Challenge Studies
4. Conclusions
Acknowledgments
Author Contributions
Conflict of Interest
EMBL Accession Number
References and Notes
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Schleiss, M.R.; McAllister, S.; Armién, A.G.; Hernandez-Alvarado, N.; Fernández-Alarcón, C.; Zabeli, J.C.; Ramaraj, T.; Crow, J.A.; McVoy, M.A. Molecular and Biological Characterization of a New Isolate of Guinea Pig Cytomegalovirus. Viruses 2014, 6, 448-475. https://doi.org/10.3390/v6020448
Schleiss MR, McAllister S, Armién AG, Hernandez-Alvarado N, Fernández-Alarcón C, Zabeli JC, Ramaraj T, Crow JA, McVoy MA. Molecular and Biological Characterization of a New Isolate of Guinea Pig Cytomegalovirus. Viruses. 2014; 6(2):448-475. https://doi.org/10.3390/v6020448
Chicago/Turabian StyleSchleiss, Mark R., Shane McAllister, Anibal G. Armién, Nelmary Hernandez-Alvarado, Claudia Fernández-Alarcón, Jason C. Zabeli, Thiruvarangan Ramaraj, John A. Crow, and Michael A. McVoy. 2014. "Molecular and Biological Characterization of a New Isolate of Guinea Pig Cytomegalovirus" Viruses 6, no. 2: 448-475. https://doi.org/10.3390/v6020448