Gene: 527 Name: Lats2 Family: Kinase Subfamily: Serine/Threonine Kinas... Accession: AB023958 GI: 7212785
Gene: 527

Gene 527
BLASTx Output


BLASTx Output. The query sequence is the Deltagen target gene.  The top "hits" are shown with
accession number, BLAST score and probability.  The alignment of the best hit is also shown.

BLAST DATE : 3/13/2003

 

                                                                         Score    E
 
 
gi|7657295|ref|NP_056586.1|  large tumor supressor 2 [Mus mu...  2003   0.0  
gi|18959200|ref|NP_055387.1|  LATSlarge tumor suppressor...  1627   0.0  
gi|7212790|dbj|BAA92381.1|  large tumor suppressor 2 [Homo s...  1538   0.0  
gi|4324436|gb|AAD16883.1|  large tumor suppressor 1 [Mus mus...   823   0.0  
 
>gi|7657295|ref|NP_056586.1| large tumor supressor 2 [Mus musculus]
 gi|7212786|dbj|BAA92380.1| warts/lats-like kinase [Mus musculus]
          Length = 1042
 
 Score = 2003 bits (5190)Expect = 0.0
 
 Identities = 987/1042 (94%)Positives = 987/1042 (94%)
 Frame = +2
 
Query: 134  MRPKTFPATTYSGNSRQRLQEIREGLKQPSKASTQGLLVGPNSDTSLDAKVLGSKDASRQ 313
            MRPKTFPATTYSGNSRQRLQEIREGLKQPSKASTQGLLVGPNSDTSLDAKVLGSKDASRQ
Sbjct: 1    MRPKTFPATTYSGNSRQRLQEIREGLKQPSKASTQGLLVGPNSDTSLDAKVLGSKDASRQ 60
 
Query: 314  QQMRATPKFGPYQKALREIRYSLLPFANESGTSAAAEVNRQMLQELVNAGCDQEMAGRAL 493
            QQMRATPKFGPYQKALREIRYSLLPFANESGTSAAAEVNRQMLQELVNAGCDQEMAGRAL
Sbjct: 61   QQMRATPKFGPYQKALREIRYSLLPFANESGTSAAAEVNRQMLQELVNAGCDQEMAGRAL 120
 
Query: 494  KQTGSRSIEAALEYISKMGYLDPRNEQIVRVIKQTSPGKGLAPTPVTRRPSFEGTGEALP 673
            KQTGSRSIEAALEYISKMGYLDPRNEQIVRVIKQTSPGKGLAPTPVTRRPSFEGTGEALP
Sbjct: 121  KQTGSRSIEAALEYISKMGYLDPRNEQIVRVIKQTSPGKGLAPTPVTRRPSFEGTGEALP 180
 
Query: 674  SYHRLGGANYEGPAALEEMPRQYLDFLFPXXXXXXXXXXXXXXPPKGYSTAVEPSAHFPG 853
            SYHRLGGANYEGPAALEEMPRQYLDFLFP              PPKGYSTAVEPSAHFPG
Sbjct: 181  SYHRLGGANYEGPAALEEMPRQYLDFLFPGAGAGTHGAQAHQHPPKGYSTAVEPSAHFPG 240
 
Query: 854  THYGRGHLLSEQPGYGVQRSSSFQNKTPPDAYSSMAKAQGGPPASLTFPAHAGLYTASHH 1033
            THYGRGHLLSEQPGYGVQRSSSFQNKTPPDAYSSMAKAQGGPPASLTFPAHAGLYTASHH
Sbjct: 241  THYGRGHLLSEQPGYGVQRSSSFQNKTPPDAYSSMAKAQGGPPASLTFPAHAGLYTASHH 300
 
Query: 1034 KPAATPPGAHPLHVLGTRGPTFTGESSAQAVLAPSRNSLNADLYELGSTVPWSAAPLARR 1213
            KPAATPPGAHPLHVLGTRGPTFTGESSAQAVLAPSRNSLNADLYELGSTVPWSAAPLARR
Sbjct: 301  KPAATPPGAHPLHVLGTRGPTFTGESSAQAVLAPSRNSLNADLYELGSTVPWSAAPLARR 360
 
Query: 1214 DSLQKQGLEASRPHVAFRAGPSRTNSFNNPQPEPSLPAPNXXXXXXXXHILHPVKSVRVL 1393
            DSLQKQGLEASRPHVAFRAGPSRTNSFNNPQPEPSLPAPN        HILHPVKSVRVL
Sbjct: 361  DSLQKQGLEASRPHVAFRAGPSRTNSFNNPQPEPSLPAPNTVTAVTAAHILHPVKSVRVL 420
 
Query: 1394 RPEPQTAVGPSHPAWVXXXXXXXXESLETKEGSAGPHPLDVDYGGSERRCXXXXXXXXXX 1573
            RPEPQTAVGPSHPAWV        ESLETKEGSAGPHPLDVDYGGSERRC          
Sbjct: 421  RPEPQTAVGPSHPAWVAAPTAPATESLETKEGSAGPHPLDVDYGGSERRCPPPPYPKHLL 480
 
Query: 1574 XXSKSEQYSVDLDSLCTSVQQSLRGGTEQDRSDKSHXXXXXXXXXXXXXQIQTSPVPVRK 1753
              SKSEQYSVDLDSLCTSVQQSLRGGTEQDRSDKSH             QIQTSPVPVRK
Sbjct: 481  LPSKSEQYSVDLDSLCTSVQQSLRGGTEQDRSDKSHKGAKGDKAGRDKKQIQTSPVPVRK 540
 
Query: 1754 NSRDEEKRESRIKSYSPYAFKFFMEQHVENVIKTYQQKVSRRLQLEQEVAKAGLCEAEQE 1933
            NSRDEEKRESRIKSYSPYAFKFFMEQHVENVIKTYQQKVSRRLQLEQEVAKAGLCEAEQE
Sbjct: 541  NSRDEEKRESRIKSYSPYAFKFFMEQHVENVIKTYQQKVSRRLQLEQEVAKAGLCEAEQE 600
 
Query: 1934 QMRKILYQKESNYNRLKRAKMDKSMFVKIKTLGIGAFGEVCLACKLDTHALYAMKTLRKK 2113
            QMRKILYQKESNYNRLKRAKMDKSMFVKIKTLGIGAFGEVCLACKLDTHALYAMKTLRKK
Sbjct: 601  QMRKILYQKESNYNRLKRAKMDKSMFVKIKTLGIGAFGEVCLACKLDTHALYAMKTLRKK 660
 
Query: 2114 DVLNRNQVAHVKAERDILAEADNEWVVKLYYSFQDKDSLYFVMDYIPGGDMMSLLIRMEV 2293
            DVLNRNQVAHVKAERDILAEADNEWVVKLYYSFQDKDSLYFVMDYIPGGDMMSLLIRMEV
Sbjct: 661  DVLNRNQVAHVKAERDILAEADNEWVVKLYYSFQDKDSLYFVMDYIPGGDMMSLLIRMEV 720
 
Query: 2294 FPEHLARFYIAELTLAIESVHKMGFIHRDIKPDNILIDLDGHIKLTDFGLCTGFRWTHNS 2473
            FPEHLARFYIAELTLAIESVHKMGFIHRDIKPDNILIDLDGHIKLTDFGLCTGFRWTHNS
Sbjct: 721  FPEHLARFYIAELTLAIESVHKMGFIHRDIKPDNILIDLDGHIKLTDFGLCTGFRWTHNS 780
 
Query: 2474 KYYQKGNHMRQDSMEPGDLWDDVSNCRCGDRLKTLEQRAQKQHQRCLAHSLVGTPNYIAP 2653
            KYYQKGNHMRQDSMEPGDLWDDVSNCRCGDRLKTLEQRAQKQHQRCLAHSLVGTPNYIAP
Sbjct: 781  KYYQKGNHMRQDSMEPGDLWDDVSNCRCGDRLKTLEQRAQKQHQRCLAHSLVGTPNYIAP 840
 
Query: 2654 EVLLRKGYTQLCDWWSVGVILFEMLVGQPPFLAPTPTETQLKVINWESTLHIPTQVRLSA 2833
            EVLLRKGYTQLCDWWSVGVILFEMLVGQPPFLAPTPTETQLKVINWESTLHIPTQVRLSA
Sbjct: 841  EVLLRKGYTQLCDWWSVGVILFEMLVGQPPFLAPTPTETQLKVINWESTLHIPTQVRLSA 900
 
Query: 2834 EARDLITKLCCAADCRLGRDGADDLKAHPFFNTIDFSRDIRKQPAPYVPTISHPMDTSNF 3013
            EARDLITKLCCAADCRLGRDGADDLKAHPFFNTIDFSRDIRKQPAPYVPTISHPMDTSNF
Sbjct: 901  EARDLITKLCCAADCRLGRDGADDLKAHPFFNTIDFSRDIRKQPAPYVPTISHPMDTSNF 960
 
Query: 3014 DPVDEESPWHEASGESAKAWDTLASPSSKHPEHAFYEFTFRRFFDDNGYPFRCPKPSEPA 3193
            DPVDEESPWHEASGESAKAWDTLASPSSKHPEHAFYEFTFRRFFDDNGYPFRCPKPSEPA
Sbjct: 961  DPVDEESPWHEASGESAKAWDTLASPSSKHPEHAFYEFTFRRFFDDNGYPFRCPKPSEPA 1020
 
Query: 3194 ESADPGDADLEGAAEGCQPVYV 3259
            ESADPGDADLEGAAEGCQPVYV
Sbjct: 1021 ESADPGDADLEGAAEGCQPVYV 1042
 
 
 

Construct

Gene:

527

 

GI Number(s):

7212785

ES Cell Line:

7148

 

 

Accession Number:

AB023958

 

 

Family:

Kinase

 

 

Subfamily:

Serine/Threonine Kinase

 

 

Species:

Mouse

 

 

 

Sequence Type:

full length cDNA

 

 

 

 

 


Sequence Length:

3460 bp

 

red underlined text = deleted in gene sequence

Sequence Deleted:

from base 2291 to base 2309

 

black text = gene sequence flanking deleted sequence



GGACAGCAATTTTTTGAAAGGAAGTTGTTCTGAAAGCATGTTTACAATTCACTGACACTG
TTGACTGTTCTCTTTAAAATAATAAGACGCTTTGAGAAGATTGTATTTATGGTAAAAGGA
AACTGGACTAACAATGAGGCCAAAGACTTTTCCTGCCACAACTTACTCTGGAAATAGCCG
GCAGCGATTGCAAGAGATTCGAGAGGGGCTGAAGCAGCCATCCAAGGCTTCCACCCAGGG
GCTGCTGGTGGGACCAAACAGTGACACTTCCCTGGATGCCAAAGTCCTGGGGAGCAAAGA
TGCCTCCAGGCAGCAGCAAATGAGAGCCACCCCGAAGTTTGGACCTTATCAAAAAGCTCT
CAGGGAAATCCGATATTCCCTCCTGCCTTTTGCCAACGAGTCAGGCACTTCGGCAGCTGC
AGAGGTGAACCGGCAGATGCTTCAGGAGTTGGTGAATGCGGGATGTGACCAGGAGATGGC
TGGCAGAGCGCTCAAGCAGACGGGCAGTAGGAGTATCGAAGCTGCCTTGGAGTACATCAG
TAAGATGGGCTACCTGGACCCCAGGAATGAGCAGATTGTGCGAGTCATCAAGCAGACCTC
CCCAGGAAAGGGCCTGGCGCCCACCCCGGTGACTCGGCGGCCCAGTTTCGAGGGCACAGG
GGAAGCACTCCCATCCTACCACCGGCTGGGTGGTGCAAACTACGAGGGCCCCGCCGCACT
GGAGGAGATGCCGCGGCAATATTTAGACTTTCTCTTCCCTGGAGCCGGAGCCGGCACCCA
CGGTGCCCAGGCTCACCAGCATCCTCCCAAAGGGTACAGCACAGCAGTAGAGCCAAGTGC
GCACTTTCCGGGCACACACTATGGTCGTGGTCATCTACTATCGGAGCAGCCTGGGTATGG
GGTGCAGCGCAGTTCCTCCTTCCAGAACAAGACGCCACCAGATGCCTATTCCAGCATGGC
CAAGGCCCAGGGTGGCCCTCCCGCCAGCCTCACCTTTCCTGCCCATGCTGGGCTGTACAC
TGCCTCGCACCACAAGCCGGCGGCTACCCCACCTGGGGCCCACCCATTACATGTGTTGGG
CACCCGGGGTCCCACGTTTACTGGCGAAAGCTCTGCACAGGCTGTGCTGGCACCGTCCAG
GAACAGCCTCAATGCTGACTTGTACGAGCTGGGCTCCACGGTGCCCTGGTCTGCAGCTCC
ACTGGCACGCCGCGACTCGCTGCAGAAGCAGGGTCTAGAAGCCTCGCGGCCGCATGTGGC
TTTTCGGGCTGGCCCCAGCAGGACCAACTCCTTCAACAACCCACAACCTGAGCCCTCACT
GCCCGCCCCCAACACGGTCACCGCCGTGACGGCCGCACACATCCTTCACCCTGTGAAGAG
CGTGCGTGTGCTGCGGCCCGAGCCCCAGACAGCCGTGGGGCCCTCGCACCCCGCCTGGGT
GGCTGCCCCCACAGCACCTGCCACTGAGAGCCTGGAGACGAAGGAGGGCAGCGCAGGCCC
ACACCCGCTGGATGTGGACTATGGCGGCTCCGAGCGCAGGTGCCCACCGCCTCCGTACCC
AAAGCACTTGCTGCTGCCCAGTAAGTCTGAGCAGTACAGCGTGGACCTGGACAGCCTGTG
CACCAGTGTGCAGCAGAGTCTGCGAGGGGGCACTGAGCAAGACAGGAGTGACAAGAGCCA
CAAAGGTGCGAAGGGAGACAAAGCTGGCAGAGACAAAAAGCAGATTCAGACCTCCCCGGT
GCCTGTCCGCAAGAATAGCAGAGATGAAGAGAAGAGAGAGTCTCGCATCAAGAGTTACTC
CCCTTATGCCTTCAAATTCTTCATGGAGCAACACGTGGAGAATGTCATCAAAACCTACCA
GCAGAAGGTCAGCCGGAGGCTACAGCTGGAGCAGGAAGTGGCCAAAGCTGGGCTCTGTGA
GGCCGAGCAGGAGCAGATGAGGAAGATCCTCTACCAGAAGGAGTCTAACTACAACCGGCT
GAAGAGGGCCAAGATGGACAAGTCCATGTTTGTGAAAATCAAGACTCTAGGCATCGGTGC
CTTTGGGGAAGTGTGCCTCGCTTGTAAGCTGGACACTCACGCTCTGTACGCCATGAAGAC
TCTCAGGAAGAAGGATGTCCTGAACCGGAATCAAGTGGCCCATGTCAAGGCTGAGAGGGA
CATCCTGGCTGAAGCAGACAATGAGTGGGTGGTCAAACTCTACTACTCCTTCCAGGACAA
GGACAGCCTGTACTTTGTGATGGACTACATACCAGGCGGGGATATGATGAGCCTGCTGAT
CAGGATGGAG
GTCTTCCCTGAGCACCTGGCCCGCTTCTACATTGCAGAGTTGACCCTGGC
CATTGAAAGTGTCCACAAGATGGGCTTTATCCACCGGGACATCAAGCCTGACAACATACT
CATCGACCTGGATGGTCATATTAAGCTGACAGATTTTGGCCTCTGCACTGGATTCAGGTG
GACTCACAATTCCAAGTACTACCAGAAAGGGAACCACATGAGACAGGACAGCATGGAGCC
CGGTGACCTCTGGGACGATGTTTCCAACTGTCGCTGTGGAGACAGGTTAAAGACCCTGGA
GCAGAGGGCGCAGAAGCAACACCAGAGGTGCCTGGCACATTCTCTTGTCGGGACACCAAA
TTACATCGCTCCGGAGGTGCTTCTCCGCAAAGGGTACACGCAGCTCTGTGACTGGTGGAG
CGTCGGTGTGATTCTCTTTGAGATGCTGGTTGGGCAGCCGCCTTTCTTGGCCCCCACCCC
CACAGAGACGCAGCTGAAGGTGATCAACTGGGAGAGCACGCTGCATATCCCTACGCAGGT
GAGGCTCAGCGCTGAGGCCCGAGACCTCATCACGAAGCTGTGCTGCGCGGCTGACTGCCG
CCTGGGCAGGGATGGGGCAGATGACCTCAAGGCACACCCGTTCTTCAACACCATCGACTT
TTCCCGTGACATCCGAAAGCAGCCTGCACCCTACGTCCCCACCATCAGCCACCCCATGGA
CACCTCCAATTTTGACCCGGTGGATGAAGAAAGCCCCTGGCACGAGGCCAGCGGAGAGAG
CGCCAAGGCCTGGGACACGCTGGCCTCCCCCAGCAGCAAGCATCCAGAGCACGCCTTCTA
TGAGTTCACCTTCCGCAGGTTCTTCGATGACAACGGCTATCCCTTCCGGTGCCCGAAGCC
CTCAGAGCCCGCAGAGAGTGCAGACCCAGGGGATGCGGACTTGGAAGGTGCGGCCGAGGG
CTGCCAGCCGGTGTACGTGTAAGCCTCAGTTAACCACAACTCGAGGAAACCCAAAATGAG
ATTTCTTTTCAGAAGACAAACTCAAGCTTAGGAATCCTTCATTTTTAGTTCTGGTAAATG
GGCAACAGGAAGAGTCAACATGATATCAAATTAGCCCTCTGAGGACCTTCACTGCATTAA
AACAGTATTTTTAAAAAATTAAAAAAAAAAAAAAAAAAAA




Targeting Vector* (genomic sequence)

    LacZ-Neo

Construct Number:  11550

Arm Length:
5':  4  kb
3':  2  kb


  * Not drawn to scale



 

5 ' PCR Reconfirmation

Gene:

527

ES Line:

7148

Gene Specific Oligo (GS):
TAAACACGACACTACGGGGCCTAGC

44901



Gel

Expected Results and Diagram

 11000bp

 10000bp

 9000bp

 8000bp

 7000bp

 6000bp

 5000bp

 4000bp

 3000bp

 2000bp

 1600bp

 1000bp

 500bp

Lane Number

Label

Expected Size (in bp)

1

1kB Ladder

 

2

44901 + NEO1 (11193)

4125

3

44901 + NEO2 (11192)

4135

4

44901 + NEO3 (2416)

4700

5

DNA Sample Control

5167

6

Positive Control

4000

 


 

3 ' PCR Reconfirmation

Gene:

527

ES Line:

7148

Gene Specific Oligo (GS):
AGCTTTGGACTCTGTCTTCATCAGC

44896



Gel

Expected Results and Diagram

 11000bp

 10000bp

 9000bp

 8000bp

 7000bp

 6000bp

 5000bp

 4000bp

 3000bp

 2000bp

 1600bp

 1000bp

 500bp

Lane Number

Label

Expected Size (in bp)

1

1kB Ladder

 

2

44896 + NEO1 (1431)

3000

3

44896 + NEO2 (1432)

2950

4

44896 + NEO3 (1170)

2250

5

DNA Sample Control

5167

6

Positive Control

2000

 


 

Legend : Genomic DNA from the recombinant ES line was assayed for homologous recombination using polymerase chain reactions (PCRs). Amplified DNA fragments were visualized by ethidium bromide staining following agarose gel electrophoresis, as shown in the gel image on the left. The test PCRs (a set of three lanes) employed the gene-specific (GS) primer, which lies outside of and adjacent to the targeting vector arm, paired in succession with one of three primers in the insertion fragment. The "DNA sample control" employed a primer pair intended to amplify a fragment from a non-targeted genomic locus. The "positive control" employed the GS primer paired with a primer at the other end of the arm. The table below the gel image lists the primers (numbered) used in each PCR and the expected product sizes, in base pairs (bp).  The diagrams to the right depict the anticipated gel image as well as the relative positions of the PCR primers.

 

 

 

3 ' Southern Blot Confirmation

Gene:

527

ES Line:

7148

Construct:

11550

Blot Number:

5107

Gene Side:

3'

 

 

MW Marker

Lane

ES Line

Enzyme

Yes

1

 

 

 

2

Parent 1

Spe I

 

3

Parent 2

Spe I

 

4

7148

Spe I

 

5

Parent 1

Bgl II

 

6

Parent 2

Bgl II

 

7

7148

Bgl II

 

Legend : Genomic DNA isolated from the indicated ES lines was digested with the indicated restriction enzymes (determined to cut outside of the construct arms).  The DNA was analyzed by Southern hybridization, probing with a radiolabeled DNA fragment that hybridizes outside of and adjacent to the construct arm.  The parent ES lines (negative controls) showed bands representing the endogenous (wild-type) Gene #527 allele.  In contrast, the ES#7148  line showed an additional band representing the targeted allele from the expected homologous recombination event.  
Note: All lanes represent data from the same blot.

 

Genotyping: Gene 527 (F2 Mice)



 

W = wild-type

H = heterozygote

K = homozygote



A) Scheme:

B) Expected Result:

 

C) Oligo Information:

Type

ID

Sequence

Multiplex Conc.

GS(E,T)

62303

CAGTGCAGAGGCCAAAATCTGTCAG

400

NEO(T)

3196

GGGTGGGATTAGATAAATGCCTGCTCT

400

GS(E1)

62302

AATCAAGTGGCCCATGTCAAGGCTG

400

 

D) Reaction Conditions:

Allele

Expected Size

Oligos to use

Targeted

540

oligo 3196 + oligo 62303

Endogenous

322

oligo 62302 + oligo 62303