Gene: 1715 Name: Prss12 Family: Protease Subfamily: Serine Protease Accession: Y13192 GI: 2113885
Gene: 1715

Gene 1715
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|6679485|ref|NP_032965.1|  proteaseserine12 neurotryps...  1518   0.0  
gi|7513606|pir||JC5759  brain-specific serine proteinase (EC...  1517   0.0  
gi|23558774|emb|CAC35028.2|  neurotrypsin [Rattus norvegicus]    1457   0.0  
gi|4506143|ref|NP_003610.1|  neurotrypsin precursor; proteas...  1167   0.0  
 
>gi|6679485|ref|NP_032965.1| proteaseserine12 neurotrypsin(motopsin) [Mus musculus]
 gi|6225760|sp|O08762|NETR_MOUSE Neurotrypsin precursor (Motopsin) (Brain-specific serine protease 3)
            (BSSP-3)
 gi|2113886|emb|CAA73646.1| neurotrypsin [Mus musculus]
 gi|2696703|dbj|BAA23986.1| serine protease [Mus musculus]
 gi|21619263|gb|AAH31429.1| proteaseserine12 neurotrypsin(motopsin) [Mus musculus]
          Length = 761
 
 Score = 1518 bits (3930)Expect = 0.0
 
 Identities = 711/752 (94%)Positives = 711/752 (94%)
 Frame = +3
 
Query: 51   VILGALSVVARADXXXXXXXXXXXXXXXXXXXXXYXXXXXXXXXXXXXXXXXXXXAAQRP 230
            VILGALSVVARAD                     Y                    AAQRP
Sbjct: 10   VILGALSVVARADPVSRSPLHRPHPSPPRSQHAHYLPSSRRPPRTPRFPLPLRIPAAQRP 69
 
Query: 231  QVLSTGHTPPTIPRRCGAGESWGNATNLGVPCLHWDEVPPFLERSPPASWAELRGQPHNF 410
            QVLSTGHTPPTIPRRCGAGESWGNATNLGVPCLHWDEVPPFLERSPPASWAELRGQPHNF
Sbjct: 70   QVLSTGHTPPTIPRRCGAGESWGNATNLGVPCLHWDEVPPFLERSPPASWAELRGQPHNF 129
 
Query: 411  CRSPDGSGRPWCFYRNAQGKVDWGYCDCGQGPALPVIRLVGGNSGHEGRVELYHAGQWGT 590
            CRSPDGSGRPWCFYRNAQGKVDWGYCDCGQGPALPVIRLVGGNSGHEGRVELYHAGQWGT
Sbjct: 130  CRSPDGSGRPWCFYRNAQGKVDWGYCDCGQGPALPVIRLVGGNSGHEGRVELYHAGQWGT 189
 
Query: 591  ICDDQWDNADADVICRQLGLSGIAKAWHQAHFGEGSGPILLDEVRCTGNELSIEQCPKSS 770
            ICDDQWDNADADVICRQLGLSGIAKAWHQAHFGEGSGPILLDEVRCTGNELSIEQCPKSS
Sbjct: 190  ICDDQWDNADADVICRQLGLSGIAKAWHQAHFGEGSGPILLDEVRCTGNELSIEQCPKSS 249
 
Query: 771  WGEHNCGHKEDAGVSCVPLTDGVIRLAGGKSTHEGRLEVYYKGQWGTVCDDGWTEMNTYV 950
            WGEHNCGHKEDAGVSCVPLTDGVIRLAGGKSTHEGRLEVYYKGQWGTVCDDGWTEMNTYV
Sbjct: 250  WGEHNCGHKEDAGVSCVPLTDGVIRLAGGKSTHEGRLEVYYKGQWGTVCDDGWTEMNTYV 309
 
Query: 951  ACRLLGFKYGKQSSVNHFDGSNRPIWLDDVSCSGKEVSFIQCSRRQWGRHDCSHREDVGL 1130
            ACRLLGFKYGKQSSVNHFDGSNRPIWLDDVSCSGKEVSFIQCSRRQWGRHDCSHREDVGL
Sbjct: 310  ACRLLGFKYGKQSSVNHFDGSNRPIWLDDVSCSGKEVSFIQCSRRQWGRHDCSHREDVGL 369
 
Query: 1131 TCYPDSDGHRLSPGFPIRLVDGENKKEGRVEVFVNGQWGTICDDGWTDKHAAVICRQLGY 1310
            TCYPDSDGHRLSPGFPIRLVDGENKKEGRVEVFVNGQWGTICDDGWTDKHAAVICRQLGY
Sbjct: 370  TCYPDSDGHRLSPGFPIRLVDGENKKEGRVEVFVNGQWGTICDDGWTDKHAAVICRQLGY 429
 
Query: 1311 KGPARARTMAYFGEGKGPIHMDNVKCTGNEKALADCVKQDIGRHNCRHSEDAGVICDYLE 1490
            KGPARARTMAYFGEGKGPIHMDNVKCTGNEKALADCVKQDIGRHNCRHSEDAGVICDYLE
Sbjct: 430  KGPARARTMAYFGEGKGPIHMDNVKCTGNEKALADCVKQDIGRHNCRHSEDAGVICDYLE 489
 
Query: 1491 KKASSSGNKEMLSSGCGLRLLHRRQKRIIGGNNSLRGAWPWQASLRLRSAHGDGRLLCGA 1670
            KKASSSGNKEMLSSGCGLRLLHRRQKRIIGGNNSLRGAWPWQASLRLRSAHGDGRLLCGA
Sbjct: 490  KKASSSGNKEMLSSGCGLRLLHRRQKRIIGGNNSLRGAWPWQASLRLRSAHGDGRLLCGA 549
 
Query: 1671 TLLSSCWVLTAAHCFKRYGNNSRSYAVRVGDYHTLVPEEFEQEIGVQQIVIHRNYRPDRS 1850
            TLLSSCWVLTAAHCFKRYGNNSRSYAVRVGDYHTLVPEEFEQEIGVQQIVIHRNYRPDRS
Sbjct: 550  TLLSSCWVLTAAHCFKRYGNNSRSYAVRVGDYHTLVPEEFEQEIGVQQIVIHRNYRPDRS 609
 
Query: 1851 DYDIALVRLQGPGEQCARLSTHVLPACLPLWRERPQKTASNCHITGWGDTGRAYSRTLQQ 2030
            DYDIALVRLQGPGEQCARLSTHVLPACLPLWRERPQKTASNCHITGWGDTGRAYSRTLQQ
Sbjct: 610  DYDIALVRLQGPGEQCARLSTHVLPACLPLWRERPQKTASNCHITGWGDTGRAYSRTLQQ 669
 
Query: 2031 AAVPLLPKRFCKERYKGLFTGRMLCAGNLQEDNRVDSCQGDSGGPLMCEKPDESWVVYGV 2210
            AAVPLLPKRFCKERYKGLFTGRMLCAGNLQEDNRVDSCQGDSGGPLMCEKPDESWVVYGV
Sbjct: 670  AAVPLLPKRFCKERYKGLFTGRMLCAGNLQEDNRVDSCQGDSGGPLMCEKPDESWVVYGV 729
 
Query: 2211 TSWGYGCGVKDTPGVYTRVPAFVPWIKSVTSL 2306
            TSWGYGCGVKDTPGVYTRVPAFVPWIKSVTSL
Sbjct: 730  TSWGYGCGVKDTPGVYTRVPAFVPWIKSVTSL 761
 
 
 

Construct

Gene:

1715

 

GI Number(s):

2113885

ES Cell Line:

6424

 

 

Accession Number:

Y13192

 

 

Family:

Protease

 

 

Subfamily:

Serine Protease

 

 

Species:

Mouse

 

 

 

Sequence Type:

full length cDNA

 

 

 

 

 


Sequence Length:

2356 bp

 

red underlined text = deleted in gene sequence

Sequence Deleted:

from base 2118 to base 2176

 

black text = gene sequence flanking deleted sequence



GGACCACACTCGGCGCCGCAGCCATGGCGCTCGCCCGCTGCGTGCTGGCTGTGATTTTAG
GGGCACTGTCTGTAGTGGCCCGCGCTGATCCGGTCTCGCGCTCTCCCCTTCACCGCCCGC
ATCCGTCCCCACCGCGTTCCCAACACGCGCACTACCTTCCCAGCTCGCGGCGGCCACCCA
GGACCCCGCGCTTCCCGCTCCCGCTGCGGATCCCCGCTGCCCAGCGCCCGCAGGTCCTCA
GCACCGGGCACACGCCCCCGACGATTCCACGCCGCTGCGGGGCAGGAGAGTCGTGGGGCA
ATGCCACCAACCTCGGCGTCCCGTGTCTACACTGGGACGAGGTGCCGCCCTTCCTGGAGC
GGTCGCCCCCGGCCAGTTGGGCTGAGCTGCGAGGGCAGCCGCACAACTTCTGCCGGAGCC
CGGATGGCTCGGGCAGACCTTGGTGCTTCTATCGGAATGCCCAGGGCAAAGTAGACTGGG
GCTACTGCGATTGTGGTCAAGGCCCGGCGTTGCCCGTCATTCGCCTTGTTGGTGGGAACA
GTGGGCATGAAGGTCGAGTGGAGCTGTACCACGCTGGCCAGTGGGGGACCATCTGTGACG
ACCAATGGGACAATGCAGACGCAGACGTCATCTGTAGGCAGCTGGGGCTCAGTGGCATTG
CCAAAGCATGGCATCAGGCACATTTTGGGGAAGGATCTGGCCCAATATTGTTGGATGAAG
TACGCTGCACCGGAAACGAGCTGTCAATTGAGCAATGTCCAAAGAGTTCCTGGGGCGAAC
ATAACTGTGGCCATAAAGAAGATGCTGGAGTGTCTTGTGTTCCTCTAACAGATGGTGTCA
TCAGACTGGCAGGAGGAAAAAGTACCCATGAAGGTCGCCTGGAGGTCTACTACAAGGGGC
AGTGGGGGACAGTCTGTGATGATGGCTGGACTGAGATGAACACATACGTGGCTTGTCGAC
TGCTGGGATTTAAATACGGCAAACAGTCCTCTGTGAACCATTTTGATGGCAGCAACAGGC
CCATATGGCTGGATGACGTCAGCTGCTCAGGAAAAGAAGTCAGCTTCATTCAGTGTTCCA
GGAGACAGTGGGGAAGGCATGACTGCAGCCATAGAGAAGATGTGGGCCTCACCTGCTATC
CTGACAGCGATGGACATAGGCTTTCTCCAGGTTTTCCCATCAGACTAGTGGATGGAGAGA
ATAAGAAGGAAGGACGAGTGGAGGTTTTTGTCAATGGCCAATGGGGAACAATCTGCGATG
ACGGATGGACCGATAAGCATGCAGCTGTGATCTGCCGGCAACTTGGCTATAAGGGTCCTG
CCAGAGCAAGGACTATGGCTTATTTTGGGGAAGGAAAAGGCCCCATCCACATGGATAATG
TGAAGTGCACAGGAAATGAGAAGGCCCTGGCTGACTGTGTCAAACAAGACATTGGAAGGC
ACAACTGCCGCCACAGTGAGGATGCAGGAGTCATCTGTGACTATTTAGAGAAGAAAGCAT
CAAGTAGTGGTAATAAAGAGATGCTCTCATCTGGATGTGGACTGAGGTTACTGCACCGTC
GGCAGAAACGGATCATTGGTGGGAACAATTCTTTAAGGGGTGCCTGGCCTTGGCAGGCTT
CCCTCAGGCTGAGGTCGGCCCATGGAGACGGCAGGCTGCTTTGTGGAGCTACCCTTCTGA
GTAGCTGCTGGGTCCTGACAGCTGCACACTGCTTCAAAAGGTACGGAAACAACTCGAGGA
GCTATGCAGTTCGAGTTGGGGATTATCATACTCTGGTCCCAGAGGAGTTTGAACAAGAAA
TAGGGGTTCAACAGATTGTGATTCACAGGAACTACAGGCCAGACAGAAGCGACTATGACA
TTGCCCTGGTTAGATTGCAAGGACCAGGGGAGCAATGTGCCAGACTAAGCACCCACGTTT
TGCCAGCCTGTTTACCTCTATGGAGAGAGAGGCCACAGAAAACAGCCTCCAACTGTCACA
TAACAGGATGGGGAGACACAGGTCGTGCCTACTCAAGAACTCTACAACAAGCTGCTGTGC
CTCTGTTACCCAAGAGGTTTTGTAAAGAGAGGTACAAGGGACTATTTACTGGGAGAATGC
TCTGTGCTGGGAACCTC
CAAGAAGACAACCGTGTGGACAGCTGCCAGGGAGACAGTGGAG
GACCACTCATGTGTGA
AAAGCCTGATGAGTCCTGGGTTGTGTATGGGGTGACTTCCTGGG
GGTATGGATGTGGAGTCAAAGACACTCCTGGAGTTTATACCAGAGTCCCCGCTTTTGTAC
CTTGGATAAAAAGTGTCACCAGTCTGTAACTTATGGAAAGCTCAAGAAATAGTAAAACAG
TAACTATTCAGTCTTC




Targeting Vector* (genomic sequence)

    LacZ-Neo

Construct Number:  11390

Arm Length:
5':  2  kb
3':  4  kb


  * Not drawn to scale



 

5 ' PCR Reconfirmation

Gene:

1715

ES Line:

6424

Gene Specific Oligo (GS):
CAGAAGTGCTGGTGCAAGTCTGAGC

43730



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

43730 + NEO1 (11193)

2125

3

43730 + NEO2 (11192)

2135

4

43730 + NEO3 (2416)

2700

5

DNA Sample Control

5167

6

Positive Control

2000

 


 

3 ' PCR Reconfirmation

Gene:

1715

ES Line:

6424

Gene Specific Oligo (GS):
TTCCTCTAGCAAGAAAGCCCATTCG

45155



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

45155 + NEO1 (1431)

5000

3

45155 + NEO2 (1432)

4950

4

45155 + NEO3 (1170)

4250

5

DNA Sample Control

5167

6

Positive Control

4000

 


 

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:

1715

ES Line:

6424

Construct:

11390

Blot Number:

5131

Gene Side:

5'

 

 

MW Marker

Lane

ES Line

Enzyme

Yes

1

 

 

 

2

Parent 1

Spe I

 

3

Parent 2

Spe I

 

4

6424

Spe 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 #1715 allele.  In contrast, the ES#6424  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 1715 (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)

57910

CATACACAACCCAGGACTCATCAG

400

NEO(T)

3196

GGGTGGGATTAGATAAATGCCTGCTCT

400

GS(E1)

57909

TGAAATCCCATTGACTAACGCCAGC

400

 

D) Reaction Conditions:

Allele

Expected Size

Oligos to use

Targeted

428

oligo 3196 + oligo 57910

Endogenous

248

oligo 57909 + oligo 57910