Gene: 731 Name: Ptpro Family: Phosphatase Subfamily: Tyrosine Phosphatase Accession: U37465 GI: 1063639
Gene: 731

Gene 731
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|10567793|gb|AAG18585.1|AF295638_1  protein tyrosine phosp...   929   0.0  
gi|8394227|ref|NP_059032.1|  receptor-type protein tyrosine ...   926   0.0  
gi|13677214|ref|NP_109592.1|  receptor-type protein tyrosine...   905   0.0  
gi|1321658|dbj|BAA08252.1|  brain-enriched membrane-associat...   890   0.0  
 
>gi|10567793|gb|AAG18585.1|AF295638_1 protein tyrosine phosphatase BK [Mus musculus]
          Length = 1226
 
 Score =  929 bits (2400)Expect = 0.0
 
 Identities = 458/490 (93%)Positives = 459/490 (93%)
 Frame = +3
 
Query: 126  KSLFAVNKTQTSVTLLWVEEGVADFFEVFCQQLGSGHNGKLQEPVAVSSHVVTISSLLPA 305
            KSLFAVNKTQTSVTLLWVEEGVADFFEV CQ+L SGHNGKLQEP AVSSHVVTISSLLPA
Sbjct: 737  KSLFAVNKTQTSVTLLWVEEGVADFFEVLCQKLASGHNGKLQEPEAVSSHVVTISSLLPA 796
 
Query: 306  TAYNCSVTSFSHDTPSVPTFIAVSTMVTEVNPNVVVISXXXXXXXXXXXXXXXXXXXXRK 485
            TAYNCSVTSFSHDTPSVPTFIAVSTMVTEVNPNVVVIS                    RK
Sbjct: 797  TAYNCSVTSFSHDTPSVPTFIAVSTMVTEVNPNVVVISVLAILSTLLIGLLLVTLVILRK 856
 
Query: 486  KHLQMARECGAGTFVNFASLEREGKLPYSWRRSVFALLTLLPSCLWTDYLLAFYINPWSK 665
            KHLQMARECGAGTFVNFASLEREGKLPYSWRRSVFALLTLLPSCLWTDYLLAFYINPWSK
Sbjct: 857  KHLQMARECGAGTFVNFASLEREGKLPYSWRRSVFALLTLLPSCLWTDYLLAFYINPWSK 916
 
Query: 666  NGLKKRKLTNPVQLDDFDSYIKDMAKDSDYKFSLQFEELKLIGLDIPHFAADLPLNRCKN 845
            NGLKKRKLTNPVQLDDFDSYIKDMAKDSDYKFSLQFEELKLIGLDIPHFAADLPLNRCKN
Sbjct: 917  NGLKKRKLTNPVQLDDFDSYIKDMAKDSDYKFSLQFEELKLIGLDIPHFAADLPLNRCKN 976
 
Query: 846  RYTNILPYDFSRVRLVSMNEEEGADYINANYIPGYNSPQEYIATQGPLPETRNDFWKMVL 1025
            RYTNILPYDFSRVRLVSMNEEEGADYINANYIPGYNSPQEYIATQGPLPETRNDFWKMVL
Sbjct: 977  RYTNILPYDFSRVRLVSMNEEEGADYINANYIPGYNSPQEYIATQGPLPETRNDFWKMVL 1036
 
Query: 1026 QQKSHIIVMLTQCNEKRRVKCDHYWPFTEEPIAYGDITVEMVSXXXXXDWASRHFRINYA 1205
            QQKSHIIVMLTQCNEKRRVKCDHYWPFTEEPIAYGDITVEMVS     DWASRHFRINYA
Sbjct: 1037 QQKSHIIVMLTQCNEKRRVKCDHYWPFTEEPIAYGDITVEMVSEEEEEDWASRHFRINYA 1096
 
Query: 1206 DEAQDVMHFNYTGWPDHGVPPANAAESILQFVFTVRQQAAKSKGPMIIHCSAGVGRTGTF 1385
            DEAQDVMHFNYT WPDHGVPPANAAESILQFVFTVRQQAAKSKG MIIHCSAGVGRTGTF
Sbjct: 1097 DEAQDVMHFNYTAWPDHGVPPANAAESILQFVFTVRQQAAKSKGTMIIHCSAGVGRTGTF 1156
 
Query: 1386 IALDRLLQHIRDHEFVDILGLVSEMRSYRMSMVQTEEQYIFIHQCVQLMWLRKKQQFCIS 1565
            IALDRLLQHIRDHEFVDILGLVSEMRSYRMSMVQTEEQYIFIHQCVQLMWLRKKQQF IS
Sbjct: 1157 IALDRLLQHIRDHEFVDILGLVSEMRSYRMSMVQTEEQYIFIHQCVQLMWLRKKQQFSIS 1216
 
Query: 1566 DVIYENVSKS 1595
            DVIYENVSKS
Sbjct: 1217 DVIYENVSKS 1226
 
 
 

Construct

Gene:

731

 

GI Number(s):

1063639

ES Cell Line:

4643

 

 

Accession Number:

U37465

 

 

Family:

Phosphatase

 

 

Subfamily:

Tyrosine Phosphatase

 

 

Species:

Mouse

 

 

 

Sequence Type:

full length cDNA

 

 

 

 

 


Sequence Length:

2760 bp

 

red underlined text = deleted in gene sequence

Sequence Deleted:

from base 324 to base 352

 

black text = gene sequence flanking deleted sequence



CAAGTGGGGAAGGACACAAAACATTCTGCCGTTGGATGTAGATGGCAGTCTTTGTGGGTG
GATGGGCAGATTCAAAAGCAATATAAAACACAGGCTCTTAGCTACCTGCCTAACCAGCCC
CTCCGAAGTCACTCTTCGCAGTGAACAAAACACAGACGTCAGTGACCCTGCTGTGGGTTG
AGGAGGGTGTTGCTGATTTCTTTGAAGTCTTCTGTCAGCAGCTCGGCTCTGGCCACAATG
GCAAACTCCAGGAGCCAGTAGCTGTGTCGTCCCACGTGGTGACCATCTCCAGCCTCCTCC
CAGCCACTGCCTACAACTGCAGT
GTCACCAGCTTCAGCCACGACACTCCCAGTGTCCCTA
CATTCATAGCTGTCTCCACAATGGTTACAGAGGTGAACCCTAACGTGGTGGTGATCTCGG
TGCTGGCCATCCTCAGCACACTTTTAATTGGACTGCTCTTAGTGACCCTTGTCATTCTCC
GAAAGAAGCACCTGCAGATGGCTAGGGAATGTGGAGCTGGCACGTTTGTCAACTTTGCAT
CCTTAGAAAGGGAAGGGAAACTCCCCTACAGTTGGCGTAGGAGTGTGTTTGCTCTCTTAA
CCCTGCTGCCTTCATGTCTTTGGACTGACTATCTTTTGGCATTTTATATTAACCCTTGGA
GTAAAAATGGCTTAAAGAAGAGGAAACTAACAAACCCCGTTCAGCTGGATGACTTCGATT
CTTACATCAAGGATATGGCCAAGGACTCGGACTATAAATTCTCTCTTCAGTTTGAGGAGT
TGAAGTTGATTGGACTGGATATTCCGCACTTTGCTGCAGATCTACCGCTGAACCGATGTA
AAAACCGCTACACAAACATCCTGCCGTATGACTTTAGCCGGGTGAGGCTAGTCTCCATGA
ACGAAGAGGAAGGAGCAGACTACATTAATGCCAACTATATTCCTGGATACAACTCACCCC
AGGAGTACATTGCCACCCAGGGTCCCCTGCCAGAAACCAGAAATGACTTCTGGAAGATGG
TCCTACAACAGAAGTCCCACATCATCGTCATGCTCACTCAATGCAATGAGAAAAGGAGGG
TAAAATGTGACCACTACTGGCCATTCACAGAAGAACCCATTGCTTATGGGGACATCACCG
TGGAGATGGTCTCTGAGGAAGAGGAGGAGGACTGGGCCAGTAGACACTTCCGGATCAACT
ATGCGGACGAAGCGCAGGACGTGATGCATTTTAACTACACAGGCTGGCCCGATCACGGTG
TGCCTCCAGCAAACGCCGCCGAGAGCATCCTGCAGTTTGTGTTCACAGTGCGACAGCAAG
CCGCCAAGAGCAAAGGGCCCATGATCATCCACTGCAGTGCGGGTGTGGGACGGACAGGAA
CCTTCATTGCCCTGGACAGGCTCCTGCAACACATTCGAGATCATGAATTTGTGGACATCT
TAGGGCTGGTATCAGAGATGCGCTCATACCGAATGTCAATGGTACAGACAGAGGAGCAGT
ACATTTTTATCCATCAGTGTGTGCAGCTGATGTGGCTGAGGAAGAAGCAACAGTTCTGCA
TCAGCGACGTCATCTACGAGAACGTCAGCAAATCCTAGTTCGGAATCTGGAGCGGTGAGG
ACATGATACCTGCGCATCCTCCCTTGCTTCCAGAGTTTTTATAGGGGCTTTTAGTCACTT
TGCTAAATCGAGTCCCTGCTGTGCAGTATATGGACAAGGAGGAGATTTACCTCCTAGAAC
CAAGAAGACCTTAAGGAGCCTACAGTATCATTTGGAGTCTTTTCACTTCTAGATGTTTGA
TGGACCAAATTCAGTAGAATTCCAGAAAGGTCACCAATGCTCCTTGAGGGGCAGGAAGCG
CAATACTTCCTGATGGCTCAGTTGGCTCTTTGCTGCTCGGGTTGGGTTGATTTTTTTTTA
AGTGCAATAATTTCTGTATAATTGTGATTTTTTTACATTCACAATTCAGAATGTTGAATT
GTTATCTGCCCACACCATCAATCAGTGCCACAGCCTTGGAAATAACCAACATCATTACGG
AGTTTTATCTCATCCCTGGAAAGGCATGGCTACACAGGAAGACGTGGTGATGAATCTCCT
TCAAAGAAACTGAGCGAGTGTGCTCCTCGCTCCAACCTTGAAGCTACGGGTCAGGAGTAG
GCAGGCAGAGGGAAGAGAGCGGTTCTACTCACCCATCAGCCAGCTTATCTTTTCCTATTT
CAAATATGAAAACCTGTGTTTCAAAGTAGAGTAGGAAAAACATTATATGTCTGACTTGTC
ATAGTGGGTTTCTTCCTTGTTGACAGTTGGGAGTTTCTTCCGTGGCTCTTTTGAACTAAT
GTCACGGTCCTTTTTGAACAGCCAGAGTTGATTCAACAGTTTTGAGTCTGACTCTGAACT
CTGAGATATCTTGGTGCCTAGCCTTCACGTTTATTTTTCTCTGTGTCCACCTGTGTACAC
CAAAACAGTTTCTCCAAGAGCTATAGGCTGTAAAAATGCACTTTCTTTCTCCACCCAGAG
GAGCTGGGAATTCAGAAGTCACGACAACAACAAACTCAGCATGTAAGGACAGGTATTGGA
CACCATCCCTTAAGAACATTGTTGAGTGTCTCTATGGATTCCGACAGGAGATTCTCTGGG
TCTTCTTTGCATTTCTGTTGTCAGAGTCATTTTAACCTGTGTAGCTAGTGGCATTATATT
CTTTGGATTTTGTATGATTAAAGTACATGATTGTGTGTTGTGACCATGAAAAAAAAAAAA




Targeting Vector* (genomic sequence)

    LacZ-Neo

Construct Number:  3780

Arm Length:
5':  2  kb
3':  2  kb


  * Not drawn to scale



 

5 ' PCR Reconfirmation

Gene:

731

ES Line:

4643

Gene Specific Oligo (GS):
CTGACCGTGGATCCAACAAATTAGC

20954



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

20954 + NEO1 (2416)

2500

3

20954 + NEO2 (2408)

2050

4

20954 + NEO3 (2750)

2025

5

DNA Sample Control

5167

6

Positive Control

2000

 


 

3 ' PCR Reconfirmation

Gene:

731

ES Line:

4643

Gene Specific Oligo (GS):
AGCCATCTGCAGGTGCTTCTTTCGG

11072



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

11072 + NEO1 (1431)

3900

3

11072 + NEO2 (1432)

3850

4

11072 + 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:

731

ES Line:

4643

Construct:

3780

Blot Number:

4469

Gene Side:

5'

 

 

MW Marker

Lane

ES Line

Enzyme

Yes

1

 

 

 

2

Parent 1

Hind III

 

3

Parent 2

Hind III

 

4

Additional 1

Hind III

 

5

4643

Hind III

 

6

Parent 1

Nco I

 

7

Parent 2

Nco I

 

8

Additional 1

Nco I

 

9

4643

Nco I

 

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 #731 allele.  In contrast, the ES#4643  line showed an additional band representing the targeted allele from the expected homologous recombination event.   "Additional" denotes a secondary line that was derived from the same targeting vector but was not used to generate mice.
Note: All lanes represent data from the same blot.

 

Genotyping: Gene 731 (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)

19211

GTATCATTACCATTGTGGAGGCAGC

400

NEO(T)

3197

GGGGATCGATCCGTCCTGTAAGTCT

400

GS(E1)

19210

CATGAGACAGAGTAACATGGAAGGG

400

 

D) Reaction Conditions:

Allele

Expected Size

Oligos to use

Targeted

600

oligo 3197 + oligo 19211

Endogenous

314

oligo 19210 + oligo 19211