313:, the template on which protein may be produced through translation. A larger number of mRNAs would express a greater amount of protein, and how many copies of mRNA are generated depends on the promoter used in the vector. The expression may be constitutive, meaning that the protein is produced constantly in the background, or it may be inducible whereby the protein is expressed only under certain condition, for example when a chemical inducer is added. These two different types of expression depend on the types of promoter and
1253:
198:
225:
cleavage sites allow for the insertion of a transgene insert. The bacteria containing the plasmids can generate millions of copies of the vector within the bacteria in hours, and the amplified vectors can be extracted from the bacteria for further manipulation. Plasmids may be used specifically as
121:
sequence that drives expression of the transgene. Simpler vectors called transcription vectors are only capable of being transcribed but not translated: they can be replicated in a target cell but not expressed, unlike expression vectors. Transcription vectors are used to amplify their insert.
298:(HACs). An artificial chromosome can carry a much larger DNA fragment than other vectors. YACs and BACs can carry a DNA fragment up to 300,000 nucleotides long. Three structural necessities of an artificial chromosome include an origin of replication, a centromere, and telomeric end sequences.
259:
are genetically engineered viruses carrying modified viral DNA or RNA that has been rendered noninfectious, but still contain viral promoters and the transgene, thus allowing for translation of the transgene through a viral promoter. However, because viral vectors frequently lack infectious
320:
Viral promoters are often used for constitutive expression in plasmids and in viral vectors because they normally force constant transcription in many cell lines and types reliably. Inducible expression depends on promoters that respond to the induction conditions: for example, the
109:) and a larger sequence that serves as the "backbone" of the vector. The purpose of a vector which transfers genetic information to another cell is typically to isolate, multiply, or express the insert in the target cell. All vectors may be used for cloning and are therefore
579:. Some of these tags may also allow for increased solubility of the target protein. The target protein is fused to the protein tag, but a protease cleavage site positioned in the polypeptide linker region between the protein and the tag allows the tag to be removed later.
365:
produced, they therefore require more components than the simpler transcription-only vectors. Expression in different host organism would require different elements, although they share similar requirements, for example a promoter for initiation of transcription, a
113:, but there are also vectors designed specially for cloning, while others may be designed specifically for other purposes, such as transcription and protein expression. Vectors designed specifically for the expression of the transgene in the target cell are called
137:. Such vectors have bacterial or viral elements which may be transferred to the non-bacterial host organism, however other vectors termed intragenic vectors have also been developed to avoid the transfer of any genetic material from an alien species.
556:
Targeting sequence: Expression vectors may include encoding for a targeting sequence in the finished protein that directs the expressed protein to a specific organelle in the cell or specific location such as the
260:
sequences, they require helper viruses or packaging lines for large-scale transfection. Viral vectors are often designed to permanently incorporate the insert into the host genome, and thus leave distinct
170:
are double-stranded extra chromosomal and generally circular DNA sequences that are capable of replication using the host cell's replication machinery. Plasmid vectors minimalistically consist of an
340:
mRNA production. These vectors are called transcription vectors. They may lack the sequences necessary for polyadenylation and termination, therefore may not be used for protein production.
529:, and an insert successfully ligated into the vector will disrupt the gene sequence, resulting in an inactive β-galactosidase. Cells containing vector with an insert may be identified using
507:: Some vectors may contain a sequence for a specific epitope that can be incorporated into the expressed protein. It allows for antibody identification of cells expressing the target protein.
541:). Cells expressing β-galactosidase (therefore does not contain an insert) appear as blue colonies. White colonies would be selected as those that may contain an insert. Other commonly used
438:
length: UTRs contain specific characteristics that may impede transcription or translation, and thus the shortest UTRs or none at all are encoded for in optimal expression vectors.
309:
of the cloned gene is a necessary component of the vector when expression of the gene is required: one gene may be amplified through transcription to generate multiple copies of
190:
conjugative - mediate DNA transfer through conjugation and therefore spread rapidly among the bacterial cells of a population; e.g., F plasmid, many R and some col plasmids.
1215:
133:. However, vectors may also have elements that allow them to be maintained in another organism such as yeast, plant or mammalian cells, and these vectors are called
470:: Promoters are used to drive the transcription of the vector's transgene as well as the other genes in the vector such as the antibiotic resistance gene. Some
457:
Modern artificially-constructed vectors contain essential components found in all vectors, and may contain other additional features found only in some vectors:
756:
Johnston C, Martin B, Fichant G, Polard P, Claverys JP (March 2014). "Bacterial transformation: distribution, shared mechanisms and divergent control".
474:
need not have a promoter for the cloned insert but it is an essential component of expression vectors so that the cloned product may be expressed.
567:: Some expression vectors include proteins or peptide sequences that allows for easier purification of the expressed protein. Examples include
708:
941:
513:: Some vectors may contain a reporter gene that allow for identification of plasmid that contains inserted DNA sequence. An example is
226:
transcription vectors and such plasmids may lack crucial sequences for protein expression. Plasmids used for protein expression, called
174:
that allows for semi-independent replication of the plasmid in the host. Plasmids are found widely in many bacteria, for example in
807:
1212:
281:
501:
allow for survival of cells that have taken up the vector in growth media containing antibiotics through antibiotic selection.
1195:
1047:
979:
935:
841:
740:
678:
1232:
1224:
272:
pattern after insertion that is detectable and indicates that the viral vector has incorporated into the host genome.
865:
del Solar, Gloria; Giraldo, Rafael; Ruiz-EchevarrĂa, MarĂa JesĂşs; Espinosa, Manuel; DĂaz-Orejas, RamĂłn (June 1998).
491:: Genetic markers for viral vectors allow for confirmation that the vector has integrated with the host genomic DNA.
645:
291:
79:
482:
640:
635:
295:
287:
1273:
1243:
102:
572:
546:
322:
141:
427:: Creates a polyadenylation tail at the end of the transcribed pre-mRNA that protects the mRNA from
441:
306:
182:
82:. Of these, the most commonly used vectors are plasmids. Common to all engineered vectors are an
576:
408:
20:
1278:
367:
269:
730:
923:
629:
478:
461:
401:
358:
231:
218:
171:
87:
83:
47:
1073:"A comparative analysis of constitutive promoters located in adeno-associated viral vectors"
404:(RBS) - follows the promoter, and promotes efficient translation of the protein of interest.
1084:
1004:
217:. These plasmid are generally non-conjugative but may have many more features, notably a "
8:
1132:"Viral promoters can initiate expression of toxin genes introduced into Escherichia coli"
1071:
Damdindorj L, Karnan S, Ota A, Hossain E, Konishi Y, Hosokawa Y, Konishi H (2014-08-29).
148:
for eukaryotic cells, although insertion of a viral vector is often called transduction.
1088:
1008:
799:
1158:
1131:
1107:
1072:
781:
498:
467:
314:
222:
193:
nonconjugative - do not mediate DNA through conjugation, e.g., many R and col plasmids.
118:
1016:
518:
481:
or other features that allow for the insertion of foreign DNA into the vector through
431:
and ensures transcriptional and translational termination: stabilizes mRNA production.
1191:
1163:
1112:
1053:
1043:
1020:
975:
931:
904:
899:
886:
866:
847:
837:
830:
773:
736:
604:
568:
558:
534:
530:
354:
349:
227:
114:
91:
27:
785:
1153:
1143:
1102:
1092:
1012:
971:
967:
894:
882:
878:
765:
176:
1236:
1219:
1097:
670:
650:
619:
471:
424:
213:
Plasmids with specially-constructed features are commonly used in laboratory for
63:
59:
55:
1257:
1229:
700:
599:
488:
464:: Necessary for the replication and maintenance of the vector in the host cell.
435:
261:
214:
206:
162:
134:
110:
51:
444:: Vectors should encode for a Kozak sequence in the mRNA, which assembles the
357:
produce proteins through the transcription of the vector's insert followed by
1267:
1057:
890:
609:
542:
510:
326:
256:
71:
851:
699:
Lodish H, Berk A, Zipursky SL, Matsudaira P, Baltimore D, Darnell J (2000).
1167:
1148:
1116:
777:
594:
395:
251:
186:. Bacterial plasmids may be conjugative/transmissible and non-conjugative:
145:
43:
1024:
908:
180:, but may also be found in a few eukaryotes, for example in yeast such as
995:
Murray, Andrew; Szostak, Jack (November 1987). "Artificial
Chromosomes".
564:
428:
235:
769:
379:
Promoter - commonly used inducible promoters are promoters derived from
864:
614:
550:
494:
391:
380:
265:
239:
286:
Artificial chromosomes are manufactured chromosomes in the context of
624:
522:
106:
129:
vectors, which contain elements necessary for their maintenance in
1252:
924:"Chapter 2 - Vectors for Gene Cloning: Plasmids and Bacteriophages"
445:
387:
264:
in the host genome after incorporating the transgene. For example,
411:
enclosed in the RBS, 8 base-pairs upstream of the AUG start codon.
336:
Some vectors are designed for transcription only, for example for
589:
504:
167:
67:
35:
420:
Eukaryote expression vectors require sequences that encode for:
836:(4th ed.). Sudbury, Mass.: Jones and Bartlett Publishers.
230:, would include elements for translation of protein, such as a
202:
197:
75:
39:
398:, which are a hybrid of both the Trp and Lac Operon promoters.
1130:
Lewin A, Mayer M, Chusainow J, Jacob D, Appel B (June 2005).
538:
140:
Insertion of a vector into the target cell is usually called
698:
333:
heat shock promoter only initiates after high temperatures.
1042:(8th ed.). Belmont, CA: Brooks/Cole Thomson Learning.
755:
362:
310:
1070:
1225:
A comparison of vectors in use for clinical gene transfer
98:
1129:
867:"Replication and Control of Circular Bacterial Plasmids"
205:
plasmid is one of the first plasmids widely used as a
1241:
370:
for translation initiation, and termination signals.
1188:
Culture of Animal Cells: A manual of basic technique
46:) used as a vehicle to artificially carry a foreign
1190:. Hoboken, New Jersey: John Wiley & Sons, Inc.
829:
1037:
962:Julin, Douglas (2014). "Artificial Chromosomes".
125:The manipulation of DNA is normally conducted on
1265:
373:
497:resistance: Vectors with antibiotic-resistance
415:
928:Gene Cloning and DNA Analysis: An Introduction
390:promoter. Other strong promoters used include
994:
525:. A multiple cloning site is located within
325:promoter only initiates transcription after
62:. A vector containing foreign DNA is termed
517:which codes for the N-terminus fragment of
871:Microbiology and Molecular Biology Reviews
827:
1157:
1147:
1106:
1096:
898:
732:Principles of Plant Genetics and Breeding
707:(4th ed.). New York: W. H. Freeman.
663:
275:
1213:Waksman Scholars introduction to vectors
1185:
966:. Springer, New York, NY. pp. 1–3.
533:by growing cells in media containing an
196:
16:Transfers genetic material between cells
1038:Solomon EP, Berg LR, Martin DW (2005).
728:
1266:
282:Artificial chromosome (disambiguation)
97:The vector itself generally carries a
66:. The four major types of vectors are
961:
921:
632:, an organism that transmits disease
13:
1178:
701:"DNA Cloning with Plasmid Vectors"
151:
14:
1290:
1206:
1017:10.1038/scientificamerican1187-62
930:(6th ed.). Wiley-Blackwell.
832:Genetics: principles and analysis
1251:
646:Bacterial artificial chromosomes
301:
292:bacterial artificial chromosomes
245:
1123:
1064:
1031:
988:
944:from the original on 2022-12-17
810:from the original on 2018-04-17
711:from the original on 2009-05-27
681:from the original on 2019-07-08
972:10.1007/978-1-4614-6436-5_91-3
955:
915:
883:10.1128/MMBR.62.2.434-464.1998
858:
821:
792:
749:
722:
692:
407:Translation initiation site -
1:
735:. John Wiley & Sons Inc.
656:
374:Prokaryotes expression vector
343:
101:sequence that consists of an
50:– usually DNA – into another
1098:10.1371/journal.pone.0106472
758:Nature Reviews. Microbiology
729:Acquaah G (16 August 2012).
641:Yeast artificial chromosomes
636:Human artificial chromosomes
477:Cloning site: This may be a
448:for translation of the mRNA.
416:Eukaryotes expression vector
296:human artificial chromosomes
288:yeast artificial chromosomes
7:
828:Hartl DL, Jones EW (1998).
583:
452:
156:
10:
1295:
1186:Freshney IR (2005-07-29).
347:
323:murine mammary tumor virus
279:
249:
160:
18:
573:glutathione-S-transferase
565:Protein purification tags
547:green fluorescent protein
521:, an enzyme that digests
268:leaves a characteristic
183:Saccharomyces cerevisiae
964:Molecular Life Sciences
577:maltose binding protein
409:Shine-Dalgarno sequence
117:, and generally have a
34:is any particle (e.g.,
1149:10.1186/1472-6750-5-19
705:Molecular Cell Biology
368:ribosomal binding site
276:Artificial chromosomes
270:retroviral integration
210:
80:artificial chromosomes
630:Vector (epidemiology)
479:multiple cloning site
462:Origin of replication
402:Ribosome binding site
232:ribosome binding site
219:multiple cloning site
200:
172:origin of replication
144:for bacterial cells,
84:origin of replication
531:blue/white selection
425:Polyadenylation tail
329:application and the
19:For other uses, see
1230:Gene Transport Unit
1089:2014PLoSO...9j6472D
1009:1987SciAm.257e..62M
997:Scientific American
770:10.1038/nrmicro3199
499:open reading frames
1235:2007-12-06 at the
1218:2008-01-18 at the
355:Expression vectors
228:expression vectors
223:restriction enzyme
211:
115:expression vectors
105:(in this case the
1274:Molecular biology
1197:978-0-471-45329-1
1136:BMC Biotechnology
1049:978-0-495-31714-2
981:978-1-4614-6436-5
937:978-1-4051-8173-0
922:Brown TA (2010).
843:978-0-7637-0489-6
804:meshb.nlm.nih.gov
742:978-1-118-31369-5
605:Expression vector
569:polyhistidine-tag
559:periplasmic space
350:Expression vector
221:" where multiple
92:selectable marker
88:multicloning site
28:molecular cloning
1286:
1256:
1255:
1247:
1201:
1172:
1171:
1161:
1151:
1127:
1121:
1120:
1110:
1100:
1068:
1062:
1061:
1035:
1029:
1028:
992:
986:
985:
959:
953:
952:
950:
949:
919:
913:
912:
902:
862:
856:
855:
835:
825:
819:
818:
816:
815:
796:
790:
789:
753:
747:
746:
726:
720:
719:
717:
716:
696:
690:
689:
687:
686:
667:
215:cloning purposes
177:Escherichia coli
48:nucleic sequence
1294:
1293:
1289:
1288:
1287:
1285:
1284:
1283:
1264:
1263:
1262:
1250:
1242:
1237:Wayback Machine
1220:Wayback Machine
1209:
1204:
1198:
1181:
1179:Further reading
1176:
1175:
1128:
1124:
1069:
1065:
1050:
1036:
1032:
993:
989:
982:
960:
956:
947:
945:
938:
920:
916:
863:
859:
844:
826:
822:
813:
811:
798:
797:
793:
754:
750:
743:
727:
723:
714:
712:
697:
693:
684:
682:
669:
668:
664:
659:
651:DNA vaccination
620:Recombinant DNA
586:
519:β-galactosidase
489:Genetic markers
472:cloning vectors
455:
418:
376:
352:
346:
304:
284:
278:
262:genetic markers
254:
248:
165:
159:
154:
152:Characteristics
135:shuttle vectors
111:cloning vectors
64:recombinant DNA
54:, where it can
24:
17:
12:
11:
5:
1292:
1282:
1281:
1276:
1261:
1260:
1240:
1239:
1227:
1222:
1208:
1207:External links
1205:
1203:
1202:
1196:
1182:
1180:
1177:
1174:
1173:
1122:
1083:(8): e106472.
1063:
1048:
1030:
987:
980:
954:
936:
914:
877:(2): 434–464.
857:
842:
820:
800:"MeSH Browser"
791:
748:
741:
721:
691:
661:
660:
658:
655:
654:
653:
648:
643:
638:
633:
627:
622:
617:
612:
607:
602:
600:Cloning vector
597:
592:
585:
582:
581:
580:
562:
554:
537:of galactose (
511:Reporter genes
508:
502:
492:
486:
475:
465:
454:
451:
450:
449:
442:Kozak sequence
439:
432:
417:
414:
413:
412:
405:
399:
375:
372:
348:Main article:
345:
342:
303:
300:
280:Main article:
277:
274:
250:Main article:
247:
244:
207:cloning vector
195:
194:
191:
163:Plasmid vector
161:Main article:
158:
155:
153:
150:
142:transformation
15:
9:
6:
4:
3:
2:
1291:
1280:
1279:Gene delivery
1277:
1275:
1272:
1271:
1269:
1259:
1254:
1249:
1248:
1245:
1238:
1234:
1231:
1228:
1226:
1223:
1221:
1217:
1214:
1211:
1210:
1199:
1193:
1189:
1184:
1183:
1169:
1165:
1160:
1155:
1150:
1145:
1141:
1137:
1133:
1126:
1118:
1114:
1109:
1104:
1099:
1094:
1090:
1086:
1082:
1078:
1074:
1067:
1059:
1055:
1051:
1045:
1041:
1034:
1026:
1022:
1018:
1014:
1010:
1006:
1002:
998:
991:
983:
977:
973:
969:
965:
958:
943:
939:
933:
929:
925:
918:
910:
906:
901:
896:
892:
888:
884:
880:
876:
872:
868:
861:
853:
849:
845:
839:
834:
833:
824:
809:
805:
801:
795:
787:
783:
779:
775:
771:
767:
764:(3): 181–96.
763:
759:
752:
744:
738:
734:
733:
725:
710:
706:
702:
695:
680:
676:
672:
666:
662:
652:
649:
647:
644:
642:
639:
637:
634:
631:
628:
626:
623:
621:
618:
616:
613:
611:
610:Hybrid vector
608:
606:
603:
601:
598:
596:
593:
591:
588:
587:
578:
574:
570:
566:
563:
560:
555:
552:
548:
544:
540:
536:
532:
528:
524:
520:
516:
512:
509:
506:
503:
500:
496:
493:
490:
487:
484:
480:
476:
473:
469:
466:
463:
460:
459:
458:
447:
443:
440:
437:
433:
430:
426:
423:
422:
421:
410:
406:
403:
400:
397:
393:
389:
385:
383:
378:
377:
371:
369:
364:
360:
356:
351:
341:
339:
334:
332:
328:
327:dexamethasone
324:
318:
316:
312:
308:
307:Transcription
302:Transcription
299:
297:
293:
289:
283:
273:
271:
267:
263:
258:
257:Viral vectors
253:
246:Viral vectors
243:
241:
237:
233:
229:
224:
220:
216:
208:
204:
199:
192:
189:
188:
187:
185:
184:
179:
178:
173:
169:
164:
149:
147:
143:
138:
136:
132:
128:
123:
120:
116:
112:
108:
104:
100:
95:
93:
89:
85:
81:
77:
73:
72:viral vectors
69:
65:
61:
57:
56:be replicated
53:
49:
45:
44:Lambda phages
41:
37:
33:
29:
22:
1187:
1139:
1135:
1125:
1080:
1076:
1066:
1039:
1033:
1003:(5): 62–68.
1000:
996:
990:
963:
957:
946:. Retrieved
927:
917:
874:
870:
860:
831:
823:
812:. Retrieved
803:
794:
761:
757:
751:
731:
724:
713:. Retrieved
704:
694:
683:. Retrieved
674:
665:
595:Viral vector
561:of bacteria.
526:
514:
456:
429:exonucleases
419:
396:Tac-Promoter
392:Trp promoter
381:
353:
337:
335:
330:
319:
305:
285:
266:retroviruses
255:
252:Viral vector
212:
181:
175:
166:
146:transfection
139:
130:
126:
124:
96:
31:
25:
359:translation
294:(BACs), or
240:stop codons
1268:Categories
948:2016-11-07
814:2018-04-16
715:2018-04-11
685:2022-04-16
675:Genome.gov
657:References
615:Minicircle
551:luciferase
495:Antibiotic
344:Expression
331:Drosophila
1058:123008833
891:1092-2172
625:Naked DNA
543:reporters
523:galactose
107:transgene
60:expressed
1233:Archived
1216:Archived
1168:15967027
1117:25170953
1077:PLOS ONE
942:Archived
852:45730915
808:Archived
786:23559881
778:24509783
709:Archived
679:Archived
671:"Vector"
584:See also
545:include
535:analogue
483:ligation
468:Promoter
453:Features
446:ribosome
434:Minimal
386:and the
338:in vitro
315:operator
290:(YACs),
168:Plasmids
157:Plasmids
119:promoter
90:, and a
68:plasmids
36:plasmids
1258:Biology
1159:1181807
1108:4149579
1085:Bibcode
1040:Biology
1025:3317814
1005:Bibcode
909:9618448
590:Plasmid
505:Epitope
361:of the
131:E. coli
127:E. coli
76:cosmids
58:and/or
40:cosmids
1244:Portal
1194:
1166:
1156:
1142:: 19.
1115:
1105:
1056:
1046:
1023:
978:
934:
907:
897:
889:
850:
840:
784:
776:
739:
575:, and
527:lacZ-α
515:lacZ-α
384:operon
317:used.
203:pBR322
103:insert
78:, and
32:vector
21:Vector
900:98921
782:S2CID
539:X-gal
311:mRNAs
236:start
1192:ISBN
1164:PMID
1113:PMID
1054:OCLC
1044:ISBN
1021:PMID
976:ISBN
932:ISBN
905:PMID
887:ISSN
848:OCLC
838:ISBN
774:PMID
737:ISBN
549:and
394:and
363:mRNA
238:and
201:The
86:, a
52:cell
30:, a
1154:PMC
1144:doi
1103:PMC
1093:doi
1013:doi
1001:257
968:doi
895:PMC
879:doi
766:doi
436:UTR
382:lac
99:DNA
26:In
1270::
1162:.
1152:.
1138:.
1134:.
1111:.
1101:.
1091:.
1079:.
1075:.
1052:.
1019:.
1011:.
999:.
974:.
940:.
926:.
903:.
893:.
885:.
875:62
873:.
869:.
846:.
806:.
802:.
780:.
772:.
762:12
760:.
703:.
677:.
673:.
571:,
388:T7
242:.
234:,
94:.
74:,
70:,
42:,
38:,
1246::
1200:.
1170:.
1146::
1140:5
1119:.
1095::
1087::
1081:9
1060:.
1027:.
1015::
1007::
984:.
970::
951:.
911:.
881::
854:.
817:.
788:.
768::
745:.
718:.
688:.
553:.
485:.
209:.
23:.
Text is available under the Creative Commons Attribution-ShareAlike License. Additional terms may apply.