Gene: 1443 Name: Acvrl1 Family: Kinase Subfamily: Serine/Threonine Kinas... Accession: L48015 GI: 1408066
Gene: 1443

Gene 1443
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|6752958|ref|NP_033742.1|  activin A receptortype II-lik...   989   0.0  
gi|15929282|gb|AAH15083.1|  activin A receptortype II-like...   986   0.0  
gi|15679968|gb|AAH14291.1|  Similar to activin A receptort...   980   0.0  
gi|2137132|pir||I48241  ALK-1 - mouse >gi|840815|emb|CAA8348...   972   0.0  
 
>gi|6752958|ref|NP_033742.1| activin A receptortype II-like 1; activin receptor-like kinase-1
            [Mus musculus]
 gi|3183063|sp|Q61288|KIR3_MOUSE Serine/threonine-protein kinase receptor R3 precursor (SKR3) (Activin
            receptor-like kinase 1) (ALK-1) (TGF-B superfamily
            receptor type I) (TSR-I)
 gi|2137120|pir||JC4337 activin receptor like kinase-1 homolog - mouse
 gi|1020393|gb|AAB03642.1| activin type IB receptor
          Length = 502
 
 Score =  989 bits (2557)Expect = 0.0
 
 Identities = 480/502 (95%)Positives = 480/502 (95%)
 Frame = +2
 
Query: 275  MTLGSFRRGLLMLSVAFGLTRGDLRRPSKLVNCTCESPHCKRPFCQGSWCTVVLVREQGR 454
            MTLGSFRRGLLMLSVAFGLTRGDLRRPSKLVNCTCESPHCKRPFCQGSWCTVVLVREQGR
Sbjct: 1    MTLGSFRRGLLMLSVAFGLTRGDLRRPSKLVNCTCESPHCKRPFCQGSWCTVVLVREQGR 60
 
Query: 455  HPQVYRGCGSLNQELCLGRPTEFLNHHCCYRSFCNHNVSLMLEATQTPSEEPEVDAHXXX 634
            HPQVYRGCGSLNQELCLGRPTEFLNHHCCYRSFCNHNVSLMLEATQTPSEEPEVDAH   
Sbjct: 61   HPQVYRGCGSLNQELCLGRPTEFLNHHCCYRSFCNHNVSLMLEATQTPSEEPEVDAHLPL 120
 
Query: 635  XXXXXXXXXXXXXXXXXXXWRVRRRQEKQRDLHSDLGESSLILKASEQADSMLGDFLDSD 814
                               WRVRRRQEKQRDLHSDLGESSLILKASEQADSMLGDFLDSD
Sbjct: 121  ILGPVLALPVLVALGALGLWRVRRRQEKQRDLHSDLGESSLILKASEQADSMLGDFLDSD 180
 
Query: 815  CTTGSGSGLPFLVQRTVARQVALVECVGKGRYGEVWRGSWHGESVAVKIFSSRDEQSWFR 994
            CTTGSGSGLPFLVQRTVARQVALVECVGKGRYGEVWRGSWHGESVAVKIFSSRDEQSWFR
Sbjct: 181  CTTGSGSGLPFLVQRTVARQVALVECVGKGRYGEVWRGSWHGESVAVKIFSSRDEQSWFR 240
 
Query: 995  ETEIYNTVLLRHDNILGFIASDMTSRNSSTQLWLITHYHEHGSLYDFLQRQTLEPQLALR 1174
            ETEIYNTVLLRHDNILGFIASDMTSRNSSTQLWLITHYHEHGSLYDFLQRQTLEPQLALR
Sbjct: 241  ETEIYNTVLLRHDNILGFIASDMTSRNSSTQLWLITHYHEHGSLYDFLQRQTLEPQLALR 300
 
Query: 1175 LAVSAACGLAHLHVEIFGTQGKPAIAHRDLKSRNVLVKSNLQCCIADLGLAVMHSQSSDY 1354
            LAVSAACGLAHLHVEIFGTQGKPAIAHRDLKSRNVLVKSNLQCCIADLGLAVMHSQSSDY
Sbjct: 301  LAVSAACGLAHLHVEIFGTQGKPAIAHRDLKSRNVLVKSNLQCCIADLGLAVMHSQSSDY 360
 
Query: 1355 LDIGNNPRVGTKRYMAPEVLDEHIRTDCFESYKWTDIWAFGLVLWEIARRTIINGIVEDY 1534
            LDIGNNPRVGTKRYMAPEVLDEHIRTDCFESYKWTDIWAFGLVLWEIARRTIINGIVEDY
Sbjct: 361  LDIGNNPRVGTKRYMAPEVLDEHIRTDCFESYKWTDIWAFGLVLWEIARRTIINGIVEDY 420
 
Query: 1535 RPPFYDMVPNDPSFEDMKKVVCVDQQTPTIPNRLAADPVLSGLAQMMRECWYPNPSARLT 1714
            RPPFYDMVPNDPSFEDMKKVVCVDQQTPTIPNRLAADPVLSGLAQMMRECWYPNPSARLT
Sbjct: 421  RPPFYDMVPNDPSFEDMKKVVCVDQQTPTIPNRLAADPVLSGLAQMMRECWYPNPSARLT 480
 
Query: 1715 ALRIKKTLQKLSHNPEKPKVIH 1780
            ALRIKKTLQKLSHNPEKPKVIH
Sbjct: 481  ALRIKKTLQKLSHNPEKPKVIH 502
 
 
 

Construct

Gene:

1443

 

GI Number(s):

1408066

ES Cell Line:

4068

 

 

Accession Number:

L48015

 

 

Family:

Kinase

 

 

Subfamily:

Serine/Threonine Kinase

 

 

Species:

Mouse

 

 

 

Sequence Type:

full length cDNA

 

 

 

 

 

Sequence Length:

1962 bp

 

red underlined text = deleted in gene sequence

Sequence Deleted:

from base 634 to base 748

 

black text = gene sequence flanking deleted sequence



GGGGAGCAGACACTCTCAGCCCAAACTCAGCTTTCTTAGCGCTGTTCCTTAGGAGTGAGC
ACAGCCCCTTCCCAGTCCCCGGAGGCCGCCGCGCCAGCCGCGCAGATCGAGACCTTTCCC
GGCCCCACAGGCCGGCTCTGGACGTGAGACCCCGGCCGCCTCCGGCAAGGAGAGGCGGGG
GTCGAGTCGCCCTGTCCAAAGGAGGAGACCTTGGTCACCACAGCTATCACGTTTTGCAGA
TCTCACCACATCTCTTCTCCTATCTCCAAGGACCATGACCTTGGGGAGCTTCAGAAGGGG
CCTTTTGATGCTGTCGGTGGCCTTTGGCCTAACCAGGGGAGACTTGCGAAGGCCTTCCAA
GCTGGTGAACTGCACTTGTGAGAGCCCACACTGCAAGAGACCATTCTGCCAGGGGTCATG
GTGCACAGTGGTGCTGGTTCGGGAGCAGGGCAGGCACCCCCAGGTCTATCGGGGCTGTGG
GAGCCTGAACCAGGAGCTCTGCTTGGGACGTCCCACAGAGTTTCTGAACCATCACTGCTG
CTATAGATCCTTCTGCAACCACAACGTGTCTCTGATGCTGGAGGCCACCCAAACTCCTTC
GGAGGAGCCAGAAGTTGATGCCCATCTGCCTCTGATCCTGGGTCCTGTGCTGGCCTTGCC
GGTCCTGGTGGCCCTGGGTGCTCTAGGCTTGTGGCGTGTCCGGCGGAGGCAGGAGAAGCA
GCGGGATTTGCACAGTGACCTGGGCGAGTCCAGTCTCATCCTGAAGGCATCTGAACAGGC
AGACAGCATGTTGGGGGACTTCCTGGACAGCGACTGTACTACGGGCAGCGGCTCGGGGCT
CCCCTTCTTGGTGCAGAGGACGGTAGCTCGGCAGGTTGCGCTGGTAGAGTGTGTGGGAAA
GGGCCGATATGGTGAGGTGTGGCGCGGTTCGTGGCATGGCGAGAGTGTGGCGGTCAAGAT
TTTCTCCTCACGAGATGAGCAGTCCTGGTTCCGGGAGACGGAGATCTACAACACAGTTCT
GCTTAGACACGACAACATCCTAGGCTTCATCGCCTCCGACATGACTTCGCGGAACTCGAG
CACGCAGCTGTGGCTCATCACCCACTACCATGAACACGGCTCCCTCTATGACTTTCTGCA
GAGGCAGACGCTGGAGCCCCAGTTGGCCCTGAGGCTAGCTGTGTCCGCGGCCTGCGGCCT
GGCGCACCTACATGTGGAGATCTTTGGCACTCAAGGCAAACCAGCCATTGCCCATCGTGA
CCTCAAGAGTCGCAATGTGCTGGTCAAGAGTAACTTGCAGTGTTGCATTGCAGACCTGGG
ACTGGCTGTGATGCACTCACAAAGCAGCGATTACCTGGATATCGGCAACAACCCCCGAGT
GGGTACCAAAAGATACATGGCACCCGAGGTGCTGGATGAGCACATCCGCACAGACTGCTT
TGAGTCGTACAAGTGGACAGACATCTGGGCCTTTGGCCTAGTGCTATGGGAGATCGCCCG
GCGGACCATCATCAATGGCATTGTGGAGGATTACAGGCCACCTTTCTATGACATGGTACC
CAATGACCCCAGTTTTGAGGACATGAAAAAGGTGGTGTGCGTTGACCAGCAGACACCCAC
CATCCCTAACCGGCTGGCTGCAGATCCGGTCCTCTCCGGGCTGGCCCAGATGATGAGAGA
GTGCTGGTACCCCAACCCCTCTGCTCGCCTCACCGCACTGCGCATAAAGAAGACATTGCA
GAAGCTCAGTCACAATCCAGAGAAGCCCAAAGTGATTCACTAGCCCAGGGCCACCAGGCT
TCCTCTGCCTAAAGTGTGTGCTGGGGCAGAAGACATAGCCTGTCTGGGTAGAGGGAGTGA
AGAGAGTGTGCACGCTGCCCTGTGTGTGCCTGCTCAGCTTGGCTCCCAGCCCATCCAGCC
AAAAATACAGCTGAGCTGAAATTCAAAAAAAAAAAAAAACCG



Targeting Vector* (genomic sequence)

    LacZ-Neo

Construct Number:  8014

Arm Length:
5':  1.5  kb
3':  2.9  kb


  * Not drawn to scale



 

5 ' PCR Reconfirmation

Gene:

1443

ES Line:

4068

Gene Specific Oligo (GS):
GACCTTGGTCACCACAGCTATCACG

29890



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

29890 + NEO1 (11193)

1425

3

29890 + NEO2 (11192)

1435

4

29890 + NEO3 (2416)

2000

5

DNA Sample Control

5167

6

Positive Control

1300

 

 

3 ' PCR Reconfirmation

Gene:

1443

ES Line:

4068

Gene Specific Oligo (GS):
AAGATTTCAAGGCCACATCCACACCC

29895



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

29895 + NEO1 (1431)

3900

3

29895 + NEO2 (1432)

3850

4

29895 + NEO3 (1170)

3150

5

DNA Sample Control

5167

6

Positive Control

2900

 

 

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.

 

 

 

5 ' Southern Blot Confirmation

Gene:

1443

ES Line:

4068

Construct:

8014

Blot Number:

3787

Gene Side:

5'

 

 

MW Marker

Lane

ES Line

Enzyme

Yes

1

 

 

 

2

Parent 1

Bam HI

 

3

Parent 2

Bam HI

 

4

4068

Bam HI

 

5

Parent 1

Eco RI

 

6

Parent 2

Eco RI

 

7

4068

Eco RI

 

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 #1443 allele.  In contrast, the ES#4068  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 1443 (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)

37575

CCTGTTCAGATGCCTTCAGGATGAG

400

NEO(T)

3196

GGGTGGGATTAGATAAATGCCTGCTCT

400

GS(E1)

37574

TCTGACTGTAAGTAGTCTGGCTCAG

400

 

D) Reaction Conditions:

Allele

Expected Size

Oligos to use

Targeted

430

oligo 3196 + oligo 37575

Endogenous

272

oligo 37574 + oligo 37575