22D4

Site of transposon insertion cloned as PstI fragment into pSP72. 
Sequenced in opposite orientations with JEP131 and JEP132,
from sites at opposing ends of the transposon, to give:

JEP131 468 bp

JEP132 465 bp

 
Composite sequence:

CNTCCGGCNC CAAGNTGGAA NTGGTNNCNG TTNGAAANTG GTNACNCTNG
GNNATCCGGG NCCATCANCC GTTANTAGAA NGGACNACCA CTCCGGCCAG
GTTANTGATG ATGGGGGCAN TCCGGCCNCC TTAATCGNCC GGGCANTGTT
CCGCCAGCGT CAGCAGTCGC TGGCGCATGT TCCNCAGCTC GCTGATGTGC
TTTTCGATCT CCGCCACCTT CTGCAGATGT TCCGCCAGGG TAAAGGTCAA
CTGGAACTCT TTGCACACCA GCCCCTGATG CAGGTACTCG ACCTGTTTTT
CCGGCGGCGT GGGTTTACCG CTGACGGTCA GCCGCGGCCC TTCCACTTCG
ATATTCAGTT CGCTCTGCCG GAAACCGGCC AGCGCCAGAG AAATACGGTA
GTGATTGTCA TCGCTTTTCT CGATGTTATA CGGCGGGAAC CCCTGGGGTT
CCGGCCGCCG GTGACAAA

NAAACCTTGA ACTGGCCAAT TTATCAAAAC CAATCCTTGA CGCAGAAGCG
GGGATAGATC GTAATTACGC ATAGGAACAA CTCCTTTCAT TAAGCGAGAT
GTGTTTATTC AGCCCCCTGA CGGCAGGCGG ATTGTTACTG CATATTTGCC
GACAGCGTTC CTGCCGGCGA AAAGGGAATT ATTTAATTTC GATACGGCGC
GGTTTTAATG TTTCCGGCAC GATGCGCTGC ATATCGATAT ACAGCAGGCC
GTTTTCCAGT TTGGCGCCTT TAATTTGAAT ATGTTCAAGC CAGCTGGAAT
TTACGCTCGA AATTACGCTC AGCGATGCCC TGATATAAAT AGGTTTTTTC
TGCCGGTTCG TTATTGTGCG CACCGCGCAC GATCAAGCAG ATTATCTTGC
GTGGTGATCT CCAGCTCCTG CTCGGNAACC CGGNCTACCG AATGGCGATG
CGATAATGAT TTTCN




BLASTX with this sequence gives hits to a family of heat shock proteins (full results here), 
with the top hit being:

>gi|22127970|ref|NP_671393.1|   (NC_004088) heat shock protein [Yersinia pestis KIM]
          Length = 168

 Score =  152 bits (385), Expect = 2e-36
 Identities = 81/112 (72%), Positives = 89/112 (79%), Gaps = 1/112 (0%)
 Frame = -3

Query: 502 IGFDKLASSRFXCHRRPEPQGFPPYNIEKSDDNHYRISLALAGFRQSELNIEVEGPRLTV 323
           IGFDKLAS+        EPQGFPPYNIEK+DDNHYRISLALAGF+QSEL+IEVEGPRLTV
Sbjct: 28  IGFDKLASTM---QGGQEPQGFPPYNIEKTDDNHYRISLALAGFKQSELDIEVEGPRLTV 84

Query: 322 SGKPTPPEKQVEYLHQGLVCKEFQLTFTLAEHLQ-KVAEIEKHISELXNMRQ 170
            GKPTP EKQVEYLHQGLV KEF LTFTLAEHL    A+ E  +  +  +RQ
Sbjct: 85  RGKPTPVEKQVEYLHQGLVRKEFSLTFTLAEHLNVDNAQFENGLLHIDLLRQ 136

 Score = 43.9 bits (102), Expect(2) = 3e-07
 Identities = 19/34 (55%), Positives = 26/34 (75%)
 Frame = -3

Query: 754 EHIQIKGAKLENGLLYIDMQRIVPETLKPRRIEI 653
           EH+ +  A+ ENGLL+ID+ R VPE L+P+RI I
Sbjct: 115 EHLNVDNAQFENGLLHIDLLRQVPEALQPQRIAI 148

 Score = 32.3 bits (72), Expect = 3.3
 Identities = 18/39 (46%), Positives = 23/39 (58%)
 Frame = -1

Query: 555 KGVVPMRNYDLSPLLRQGLVLINWPVQGXFVTGGRNPRG 439
           K +  MRNYDLSPLLRQ    I +      + GG+ P+G
Sbjct: 10  KELTYMRNYDLSPLLRQ---WIGFDKLASTMQGGQEPQG 45

 Score = 31.6 bits (70), Expect(2) = 3e-07
 Identities = 18/59 (30%), Positives = 26/59 (43%)
 Frame = -2

Query: 932 ENHYRIAIR*XGXXXXXXXXXRKIICLIVRGAHNNEPAEKTYLYQGIAERNFERKFQLA 756
           +NHYRI++   G          +   L VRG       +  YL+QG+  + F   F LA
Sbjct: 56  DNHYRISLALAGFKQSELDIEVEGPRLTVRGKPTPVEKQVEYLHQGLVRKEFSLTFTLA 114

The transposon is thus inserted within a gene encoding a homologue of a heat-shock protein of the ibpB family. The E. coli ibpB, protect enzymes from inactivation by heat and oxidants (see abstract by Kitagawa et al.). The Colibri entry for ibpB shows that ibpB is downstream of a second heat shock protein, ibpA, surrounded by genes of unknown function. See also the entry for ibpB at EcoGene.