Mike's Moss Resources

Physcomitrium patens links (formerly Physcomitrella patens)

Methods (Bezanilla Lab)
Methods (Hasebe)
Methods (Quatrano & Cove)
CoGe Physcomitrella genome resources
Physcomitrella eFP Browser
Phytozome (comparative plant genomics)

Mike's Moss Transformation Protocol

ApE - A Plasmid Editor (for seq files)

pMP series vectors

Gateway Destination vectors for over-expression/inducible expression
pMP1155 (pUK-Pp108B-nptII-Ubi:Gateway:ocs)         Seq
pMP1228 (pUK-Pp108B-nptII-Ubix:Gateway:ocs)(no intron)    Seq
pMP1180 (pUK-Pp108B-nptII-GmHsp17.3B:Gateway:e9)    Seq
pMP1432 (pUK-Pp108B-Gent-GmHsp17.3B:Gateway:e9)    Seq


Moss selectable marker plasmids(derived from pBNRF--a few more RE sites are unique)

G418 (20 µg/ml) and Hyg (15 µg/ml) selections work better than others. Zeocin (25 µg/ml) also works fairly well, but seems to induce some unrelated growth phenotypes occasionally (via DNA damage?). Gentamicin (50-100 µg/ml) selection can tricky, but we have had good luck lately. We use Gentamicin Sulfate from Chem Impex (cat 00149). Kanamycin (100 µg/ml) takes a while to kill sensitive moss cells, so we have only used it in dual selections with another antibiotic.

pMP1109 (pBiLox+35S:nptII:camv) G418/Kanamycin-res*.    Seq
pMP1159 (pBiLox+2x35S:aph(4)-Ia:camv) Hygromycin-res.    Seq
pMP1169 (pBiLox+35S:Zeo:camv) Zeocin-res.    Seq
pMP1119 (pBiLox+35S:aacC1:camv) Gentamycin/G418-res*.    Seq

*Expression of aacC1 in moss confers resistance to gentamicin and G418 but not kanamycin, while nptII expression confers resistance to G418 and kanamycin but not gentamicin.

Gateway Destination vectors for over-expression with tags

pMP1300 (pUK-Pp108B-nptII-Ubi:Gateway:GUS:ocs)    Seq
pMP1367 (pUK-Pp108B-nptII-Ubi:Gateway:TAP:ocs)    Seq
pMP1302 (pUK-Pp108B-nptII-Ubi:Gateway:TAP2:ocs)    Seq
pMP1298 (pUK-Pp108B-nptII-Ubi:Gateway:GFPhdel:ocs)    Seq
pMP1382 (pUK-Pp108B-nptII-Ubi:Gateway:GFP-His:ocs)    Seq
pMP1379 (pUK-Pp108B-nptII-Ubi:Gateway:YFP-HA:ocs)    Seq
pMP1574 (pUK-Pp108B-nptII-Ubi:Gateway:CFP-HA:ocs)    Seq
pMP1472 (pUK-Pp108B-nptII-Ubi:Gateway:YFPnls-HA:ocs)    Seq
pMP1442 (pUK-Pp108B-nptII-Ubi:Gateway:mVenus-HA:ocs)    Seq
pMP1474 (pUK-Pp108B-nptII-Ubi:Gateway:mVenusnls-HA:ocs)    Seq
pMP1445 (pUK-Pp108B-nptII-Ubi:Gateway:DsRED2-HA:ocs)    Seq
pMP1475 (pUK-Pp108B-nptII-Ubi:Gateway:DsRED2nls-HA:ocs)    Seq
pMP1377 (pUK-Pp108B-nptII-Ubi:Gateway:4xMyc:nos)    Seq
pMP1373 (pUK-Pp108B-nptII-Ubi:Gateway:1xMyc:ocs)    Seq
pMP1369 (pUK-Pp108B-nptII-Ubi:Gateway:HA:ocs)    Seq
pMP1371 (pUK-Pp108B-nptII-Ubi:Gateway:FLAG:ocs)    Seq
pMP1375 (pUK-Pp108B-nptII-Ubi:Gateway:AcV5:ocs)    Seq
pMP1575 (pUK-Pp108B-nptII-Ubi:Gateway:GRlbd:ocs)    Seq
pMP1578 (pUK-Pp108B-nptII-Ubi:Gateway:ESR1lbd:ocs)    Seq
pMP1335 (pUK-Pp108B-nptII-Ubi:GFP6:Gateway:ocs)    Seq
pMP1361 (pUK-Pp108B-nptII-Ubi: YFP:Gateway:ocs)    Seq
pMP1363 (pUK-Pp108B-nptII-Ubi:TAP2:Gateway:ocs)    Seq
pMP1359 (pUK-Pp108B-nptII-Ubi:1xMyc:Gateway:ocs)    Seq
pMP1355 (pUK-Pp108B-nptII-Ubi:HA:Gateway:ocs)    Seq
pMP1357 (pUK-Pp108B-nptII-Ubi:FLAG:Gateway:ocs)    Seq
pMP1365 (pUK-Pp108B-nptII-Ubi:AcV5:Gateway:ocs)    Seq


Physcomitrella neutral target loci plasmids

pMP1141 (pGEM+Pp108A) hydrolase target    Seq
pMP1033 (pUK+Pp108B) adjacent intergenic region target    Seq
Endogenous Pp108 target locus    Seq